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  • Biochemistry and Biotechnology  (670)
  • AERODYNAMICS
  • ddc:330
  • 2000-2004
  • 1995-1999  (730)
  • 1985-1989
  • 1980-1984
  • 1945-1949
  • 1925-1929
  • 1999  (33)
  • 1996  (697)
Collection
Language
Years
  • 2000-2004
  • 1995-1999  (730)
  • 1985-1989
  • 1980-1984
  • 1945-1949
  • +
Year
  • 1
    Publication Date: 2019-07-13
    Description: The implementation of a two-equation k-omega turbulence model into the NPARC flow solver is described. Motivation for the selection of this model is given, major code modifications are outlined, new imputs to the code are described, and results are presented for several validation cases: an incompressible flow over a smooth flat plate, a subsonic diffuser flow, and a shock-induced separated flow. Comparison of results with the k-epsilon model indicate that the k-omega model predicts simple flows equally well whereas, for adverse pressure gradient flows, the k-omega model outperforms the other turbulence models in NPARC.
    Keywords: AERODYNAMICS
    Type: NASA-TM-107080 , NAS 1.15:107080 , E-9955 , AIAA PAPER 96-0383 , NIPS-96-08118 , Aerospace Sciences Meeting and Exhibit; Jan 15, 1996 - Jan 18, 1996; Reno, NV; United States
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  • 2
    Publication Date: 2019-07-13
    Description: An approach for solving the compressible Euler and Navier-Stokes equations upon meshes composed of nearly arbitrary polyhedra is described. Each polyhedron is constructed from an arbitrary number of triangular and quadrilateral face elements, allowing the unified treatment of tetrahedral, prismatic, pyramidal, and hexahedral cells, as well the general cut cells produced by Cartesian mesh approaches. The basics behind the numerical approach and the resulting data structures are described. The accuracy of the mixed volume grid approach is assessed by performing a grid refinement study upon a series of hexahedral, tetrahedral, prismatic, and Cartesian meshes for an analytic inviscid problem. A series of laminar validation cases are made, comparing the results upon differing grid topologies to each other, to theory, and experimental data. A computation upon a prismatic/tetrahedral mesh is made simulating the laminar flow over a wall/cylinder combination.
    Keywords: AERODYNAMICS
    Type: NASA-TM-107135 , NAS 1.15:107135 , AIAA PAPER 96-0762 , E-10065 , NIPS-96-07909 , Aerospace Sciences Meeting and Exhibit; Jan 15, 1996 - Jan 18, 1996; Reno, NV; United States
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  • 3
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    Bonn : Friedrich-Ebert-Stiftung | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-11-19
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
    Type: report , doc-type:report
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  • 4
    Publication Date: 2016-04-28
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
    Type: workingpaper , doc-type:workingPaper
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  • 5
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    Bonn : Dietz | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 6
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    San Francisco : Sierra Club Books | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: English
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  • 7
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 8
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
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    Language: German
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  • 9
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 10
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    Berlin : Birkhäuser | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2014-08-15
    Description: Ökoeffizienz ist die Grundlage eines zukunftsfähigen Managements, dessen Ziel es ist, Ökonomie und Ökologie zu vereinen. Ernst Ulrich von Weizsäcker und Jan-Dirk Seiler-Hausmann zeigen, daß diese Kombination den Unternehmen in Zukunft sogar mehr Gewinn bringen kann als herkömmliche Unternehmensführung.
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 11
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    Bonn : Bündnis 90/Die Grünen | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 12
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    Cheltenham : Elgar | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-04-30
    Keywords: ddc:330
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  • 13
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    Wiesbaden : Gabler | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-04-30
    Keywords: ddc:330
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    Language: German
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  • 14
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2014-08-15
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 15
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-04-18
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 16
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
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  • 17
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2014-08-15
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  • 18
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-04-30
    Keywords: ddc:330
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    Language: German
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  • 19
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    Leipzig : Brockhaus | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2014-08-15
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 20
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    Salzburg : Eigenverl. | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
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    Language: German
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  • 21
    Publication Date: 2020-10-13
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 22
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2014-08-15
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  • 23
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2014-08-15
    Keywords: ddc:330
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  • 24
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-04-30
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  • 25
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    Aachen : Kersting | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-03-26
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 26
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    Berlin : Schmidt | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
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  • 27
    Publication Date: 2018-04-18
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 28
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
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  • 29
    Publication Date: 2014-08-15
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 30
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    Berlin : Birkhäuser | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
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    Language: German
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  • 31
    Publication Date: 2016-04-28
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
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  • 32
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-11-21
    Description: Statisticians avoid getting involved in data analysis, leaving data users on their own in interpreting the results of their work. This is particularly unfortunate in a new area of applied statistics such as environmental accounting with which few are really familiar. Earlier this year data producers and users explored, in a national seminar, possible policy applications of the results of a "green accounting" project in the Philippines. The main findings of the author's contribution to the seminar, on which the present paper is based, are that environmental accounts: (1) present evidence of sustainable economic performance in the country during the relatively short-time period of 1988–1994; (2) provide information for environmental cost internalization; (3) may guide investment to environmentally sound production processes; (4) help to specify and monitor policies of natural wealth conservation, distribution and management; and (5) reveal major data gaps. The paper concludes that environmental accounts help to assess the sustainability of economic growth in terms of broadly defined capital maintenance. The sustainability of development, however, would have to be measured by alternative or supplementary physical indicators linked to quantifiable standards or targets.
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
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  • 33
    Publication Date: 2018-04-30
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    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
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  • 34
    Publication Date: 2018-04-30
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  • 35
    Publication Date: 2018-04-30
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  • 36
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
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  • 37
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    Cheltenham : Elgar | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
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  • 38
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
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  • 39
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    Berlin : Birkhäuser | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-04-11
    Keywords: ddc:330
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  • 40
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
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  • 41
    Publication Date: 2016-04-28
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  • 42
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    Bad Neuenahr-Ahrweiler : Europ. Academy for the Study of Consequences of Scientific and Technological Advance | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
    Keywords: ddc:330
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  • 43
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    Unna : Stadtwerke Unna | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2016-04-28
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  • 44
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-04-30
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  • 45
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
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  • 46
    Publication Date: 2016-04-28
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  • 47
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2014-08-15
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  • 48
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  • 49
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
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  • 50
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    München : Hampp | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2014-08-15
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  • 51
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
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  • 52
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
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  • 53
    Publication Date: 2018-11-19
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
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  • 54
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-11-21
    Description: This paper compares the one-sector neoclassical (Solow) growth model with a neo-Austrian growth model. The solutions of the neoclassical growth model in terms of the golden rule of accumulation and the Ramsey rule are well known, and these conditions are compared with the outcomes of the neo-Austrian growth model, which we derive using a basic three-process model.
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  • 55
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    Gelsenkirchen : Inst. Arbeit und Technik | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2018-11-19
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  • 56
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2022-02-18
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: English
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2022-02-18
    Description: Dieser Beitrag sucht nach Argumenten für und gegen die Langlebigkeit von Produkten aus ökologischer und ökonomischer Sicht, um herauszufinden, wie das Konzept der Produktlanglebigkeit so umgesetzt werden könnte, daß existierende Nachteile minimiert und mögliche Vorteile maximiert werden. In Kapitel 2 wird zunächst die Forderung nach langlebigen Produkten von ökologischer Seite beleuchtet. Neben der Begründung für das Konzept der Produktlanglebigkeit werden Problembereiche erfaßt, die gegen langlebige Produkte sprechen. In Kapitel 3 werden daraufhin Erklärungsansätze für das Verhalten von wirtschaftlichen Akteuren in Bezug auf die Langlebigkeit von Produkten hergeleitet. In Kapitel 4 wird schließlich die Synthese zwischen den Ergebnissen des ökonomischen sowie des ökologischen Kapitels hergestellt. Es werden dort konkrete Lösungsansätze vorgeschlagen, mit deren Hilfe eine optimale Umsetzung des Konzeptes der Produktlanglebigkeit erfolgen kann.
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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    Publication Date: 2022-02-18
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2022-02-18
    Keywords: ddc:330
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    Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie | Wuppertal : Wuppertal Institut für Klima, Umwelt, Energie
    Publication Date: 2022-02-18
    Keywords: ddc:330
    Repository Name: Wuppertal Institut für Klima, Umwelt, Energie
    Language: German
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  • 61
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 204-216 
    ISSN: 0006-3592
    Keywords: expanded bed adsorption ; bakers' yeast ; G6PDH ; STREAMLINE ion exchange adsorbents ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The use of expanded beds of STREAMLINE ion exchange adsorbents for the direct extraction of an intracellular enzyme glucose-6-phosphate dehydrogenase (G6PDH) from unclarified yeast cell homogenates has been investigated. It has been demonstrated that such crude feedstocks can be applied to the bed without prior clarification steps. The purification of G6PDH from an unclarified yeast homogenate was chosen as a model system containing the typical features of a direct extraction technique. Optimal conditions for the purification were determined in small scale, packed bed experiments conducted with clarified homogenates. Results from these experiments were used to develop a preparative scale separation of G6PDH in a STREAMLINE 50 EBA apparatus. The use of an on-line rotameter for measuring and controlling the height of the expanded bed when operated in highly turbid feedstocks was demonstrated. STREAMLINE DEAE has been shown to be successful in achieving isolation of G6PDH from an unclarified homogenate with a purification factor of 12 and yield of 98% in a single step process. This ion exchange adsorbent is readily cleaned using simple cleaning-in-place procedures without affecting either adsorption or the bed expansion properties of the adsorbent after many cycles of operation. The ability of combining clarification, capture, and purification in a single step will greatly simplify downstream processing flowsheets and reduce the costs of protein purification. © 1996 John Wiley & Sons, Inc.
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  • 62
    Electronic Resource
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    Biotechnology and Bioengineering 49 (1996), S. 259-265 
    ISSN: 0006-3592
    Keywords: hepatocytes ; lactose-derivatized polystyrene ; polystyrene ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hepatocytes isolated from male Fisher 344VF rats were cultured on two substrates, collagen I and a lactose-derivatized polystyrene (PS-lactose), to compare morphological and functional differences. Hepatocyte morphology changed dramatically depending upon the substrate, shown through actin cytoskeletal staining and scanning electron microscopy. Functional assays performed included albumin secretion, reduced glutathione content, UDP-glucuronosyl transferase, and cytochrome P4501A1 activity. The presence of dexamethasone and dimethylsulfoxide (DMSO) in the media was required for the maintenance of several differentiated functions for cells cultured on collagen. In general, cells cultured on the PS-lactose substrate showed a much slower loss of function over the same period of time. The maintenance of differentiated function of cells on PS-lactose was enhanced with the addition of dexamethasone and DMSO. This is the first report of a culture system in which hepatocytes, cultured on a polymer substrate without additional protein coatings or media additives, have been able to maintain differentiated functions for up to 1 week. © 1996 John Wiley & Sons, Inc.
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  • 63
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 290-299 
    ISSN: 0006-3592
    Keywords: proteins, modified ; partitioning in aqueous system ; thaumatin ; β-lactoglobulin ; BSA ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Relatively conservative modifications of three proteins were carried out to alter their surface properties. The protein properties modified were hydrophobicity and charge. This was done by acylation of amino groups with anhydrides. For the hydrophobic modification experiments, two proteins (β-lactoglobulin and bovine serum albumin [BSA]) and four anhydrides (hexanoic, butyric, succinic, acetic) were used. For the modification of surface charge the protein thaumatin was selected and various proportions of the free amino groups were blocked with acetic anhydride to give a series of proteins with differing isoelectric points. Detailed characterization and purification of selected modified proteins was carried out including molecular weight measurements and conformational analysis. The criteria used for selecting the modified proteins for subsequent investigation of their partitioning in aqueous two-phase systems (ATPS) is described. With a judicious choice of starting material it was found that limited chemical modifications to proteins could effectively alter surface hydrophobicity or charge almost independently, with little effect on other molecular properties. It appears, however, that the method for chemical modification and the reaction conditions must also be carefully controlled. © 1996 John Wiley & Sons, Inc.
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  • 64
    Electronic Resource
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    Biotechnology and Bioengineering 49 (1996), S. 309-315 
    ISSN: 0006-3592
    Keywords: surface charge ; proteins, modified ; partitioning in aqueous system ; thaumatin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A series of charge-modified thaumatins with different values of surface charge were partitioned in aqueous two-phase systems (ATPS) to study the effect of surface charge as a single property on partitioning. Electrophoretic mobility of the proteins in titration curves was used as a measure of surface charge. Four modified proteins derived from thaumatin with the following values of isoelectric point: 8.70, 8.15, 5.60, and 4.50 were used for partitioning. The resolution of the systems in terms of protein surface charge was calculated. Partitioning of modified thaumatins in PEG 4000/dextran systems with phosphate buffer, Tris buffer, NaCl, KCl, and sulfate salts was carried out. Among the sulfate salts tested, the addition of 50 mM Li2SO4 to the system buffered with phosphate gave the highest value of resolution for differences in surface protein charge (RSPC). It shows a decrease in the value of K (partition coefficient) with an increase in the protein's charge. The addition of 100 mM KCl to the system promoted the opposite effect on the RSPC value. Charge-modified proteins were partitioned in PEG/salt systems to investigate the ability of these systems for resolving differences in surface charge. The PEG/citrate system seemed to have almost no ability for resolving proteins on the basis of surface charge differences; PEG/phosphate systems had some capability for resolving differently charged proteins. The more negative proteins tended to have higher values of K than the more positively charged fractions. The use of charge-modified proteins allowed the investigation of the effect of protein surface charge on partitioning in aqueous two-phase systems independently from other protein parameters as they were prepared from a common parent protein thaumatin. This technique provides an interesting novel tool to investigate the effect of protein surface charge on partitioning in ATPS taking protein charge as an independent parameter. © 1996 John Wiley & Sons, Inc.
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  • 65
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    Biotechnology and Bioengineering 49 (1996), S. 348-354 
    ISSN: 0006-3592
    Keywords: oxygenator ; NMR spectroscopy ; organ perfusion ; mammalian cell culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A compact, reusable membrane oxygenator has been constructed for the perfusion of cultured cells and isolated organs. While the oxygenator was designed to be compatible with nuclear magnetic resonance (NMR) spectroscopy studies, it can also be used for any experiment which requires warming and oxygenation of perfusates. For the NMR studies, the oxygenator can be positioned at the opening of the magnet bore which allows oxygenation and warming of the perfusate immediately prior to delivery to the tissue, therefore eliminating problems with heat or oxygen loss which may occur with the long perfusion lines. © 1996 John Wiley & Sons, Inc.
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  • 66
    ISSN: 0006-3592
    Keywords: c-fos protein ; endothelium ; hemodynamics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The c-fos protein belongs to a family of transcriptional cofactors that can complex with proteins of the Jun family and activate mRNA transcription from gene promoters containing an activator protein 1 (AP-1) binding element. The shear stress inducibility of the c-fos protein was studied in human and animal cell lines of vastly different origins. Primary human umbilical vein endothelial cells (HUVEC), bovine aortic endothelial cells (BAEC, passage 2-14), HeLa cells, and Chinese hamster ovary (CHO) cells were subjected to steady laminar shear stress using a parallel plate flow apparatus. After 1 h of flow exposure at 25 dyn/cm2, the c-fos levels in nuclei of shear stress HUVEC, BAEC, HeLa, and CHO were 5.4 ± 2.0 (n = 3), 2.25 ± 1.38 (n = 6), 2.14 ± 0.07 (n = 8), 1.92 ± 0.58 (n = 2) times higher, respectively, than in matched stationary controls. Flow exposure at 4 dyn/cm2 caused no enhancement of c-fos levels in any of the cell lines tested, but caused significant reduction in c-fos expression in the HeLa cells. The c-fos induction by shear stress could be blocked by pharmacological agents. For example, the flow induction of the c-fos protein levels was blocked by 50% with the preincubation of HUVEC with a protein kinase C inhibitor, H7 (10 μM) and blocked completely in HeLa cells preincubated with the phospholipase C inhibitor, neomycin (5 mM). The minimum time of shear stress exposure required to induce the c-fos protein expression in HeLa cells was found to be as low as 1 min. By Northern analysis, the c-fos mRNA levels were found to be elevated in BAEC, CHO, and HeLa cells exposed to 25 dyn/cm2 for 30 min. These studies indicate that c-fos induction is a consistent genetic response in a variety of mammalian cells that may alter cellular phenotype in mechanical environments. © 1996 John Wiley & Sons, Inc.
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  • 67
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    Biotechnology and Bioengineering 49 (1996), S. 412-420 
    ISSN: 0006-3592
    Keywords: Amycolatopsis orientalis ; vancomycin production ; chemostat culture ; phosphate inhibition ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Production of the glycopeptide antibiotic vancomycin by two Amycolatopsis orientalis strains was examined in batch shake flask culture in a semidefined medium with peptone as the nitrogen source. Different growth and production profiles were observed with the two strains; specific production (Yp/x) was threefold higher with strain ATCC 19795 than with strain NCIMB 12945. A defined medium with amino acids as the nitrogen source was developed by use of the Plackett-Burman statistical screening method. This technique identified certain amino acids (glycine, phenylalanine, tyrosine, and arginine) that gave significant increased specific production, whereas phosphate was identified as inhibitory for high specific vancomycin production. Experiments made with the improved medium and strain ATCC 19795 showed that vancomycin production kinetics were either growth dissociated or growth associated, depending on the amino acid concentration. In chemostat culture at a constant dilution rate (0.087 h-1), specific vancomycin production rate (qvancomycin) decreased linearly as the medium phosphate concentration was increased from 2 to 8 mM. In both phosphate and glucose limited chemostats, qvancomycin was a function of specific growth rate; the maximum value was observed at D = 0.087 h-1 (52% of the maximum specific growth rate). Under phosphate limited growth conditions, qvancomycin was threefold higher (0.37 mg/g dry weight/h) than under glucose limitation (0.12 mg/g dry weight/h). © 1996 John Wiley & Sons, Inc.
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  • 68
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    Biotechnology and Bioengineering 50 (1996), S. 36-48 
    ISSN: 0006-3592
    Keywords: insect cell culture ; Sf-9 cells ; respiration ; bioreactor ; on-line monitoring ; baculovirus expression vector system ; recombinant proteins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Respiration rates in Spodoptera frugiperda (Sf-9) cell bioreactor cultures were successfully measured on-line using two methods: The O2 uptake rate (OUR) was determined using gas phase pO2 values imposed by a dissolved oxygen controller and the CO2 evolution rate (CER) was measured using an infrared detector. The measurement methods were accurate, reliable, and relatively inexpensive. The CER was routinely determined in bioreactor cultures used for the production of several recombinant proteins. Simple linear relationships between viable cell densities and both OUR and CER in exponentially growing cultures were used to predict viable cell density. Respiration measurements were also used to follow the progress of baculoviral infections in Sf-9 cultures. Infection led to increases in volumetric and per-cell respiration rates. The relationships between respiration and several other culture parameters, including viable cell density, cell protein, cell volume, glucose consumption, lactate production, viral titer, and recombinant β-galactosidase accumulation, were examined. The extent of the increase in CER following infection and the time postinfection at which maximum CER was attained were negatively correlated with the multiplicity of infection (MOI) at multiplicities below the level required to infect all the cells in a culture. Delays in the respiration peak related to the MOI employed were correlated with delays in the peak in recombinant protein accumulation. DO levels in the range 5-100% did not exert any major effects on viable cell densities, CER, or product titer in cultures infected with a baculovirus expressing recombinant β-galactosidase. © 1996 John Wiley & Sons, Inc.
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  • 69
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    Biotechnology and Bioengineering 50 (1996) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 70
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    Biotechnology and Bioengineering 50 (1996), S. 169-183 
    ISSN: 0006-3592
    Keywords: liposomes ; biotin ; aggregation kinetics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The aggregation of biotinylated phospholipid vesicles (liposomes) cross-linked by antibiotin IgG was studied experimentally and theoretically. The liposomes were either low density liposomes that contained 0.4 mol% biotinylated phospholipid (≈100 exposed biotin molecules per liposome), or high density liposomes that contained 2.7 mol% biotinylated phospholipid (≈1000 exposed biotin molecules per liposome). The solution turbidity and mean particle size measured by quasi-elastic light scattering (QLS) were monitored throughout the aggregation. Three different lots of antibiotin antibodies, each with different association constants and binding heterogeneities, were used. The antibody binding characteristics affected the aggregation rates. The aggregation kinetics were analyzed using a model based on the Smoluchowski theory of aggregation, fractal concepts of aggregate microstructure, and Rayleigh and Mie light scattering theory. The experimental conditions of liposome concentration, protein concentration, and ligand density under which aggregation occurred correlated well with calculated sticking probabilities based on isotherms describing the adsorption of antibiotin antibody to the liposomes. These results are compared with prior observations made when avidin was used as the cross-linking protein. © 1996 John Wiley & Sons, Inc.
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  • 71
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    Biotechnology and Bioengineering 50 (1996) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 72
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    Biotechnology and Bioengineering 50 (1996), S. 211-216 
    ISSN: 0006-3592
    Keywords: microgravity ; bioprocessing ; sedimentation ; turbulence ; collagenase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of a quiescent microgravity fluid environment on the activity of collagenase directed at demineralized bone fragments was investigated over a period of 10 days. Enzyme treatment resulted in greater mass loss in microgravity, with nearly three times the loss of mass during Space Shuttle mission STS-62 compared to the stationary ground control. Clinorotation enhanced the loss of mass relative to a stationary control, but this increase was still significantly less than the increase with exposure to microgravity. This suggests the detrimental influence of turbulence on the enzyme function and the benefit of using microgravity to provide both low turbulence and uniformity of unequally dense materials within the reaction chamber. The results are considered for their general applicability to a variety of bioprocessing applications that may be enhanced in microgravity. © 1996 John Wiley & Sons, Inc.
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  • 73
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    Biotechnology and Bioengineering 50 (1996), S. 430-437 
    ISSN: 0006-3592
    Keywords: cartilage ; tissue regeneration ; chondrocytes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In the last 5 to 10 years, tissue engineering has revolutionized the way in which medical researchers and clinicians are thinking of and, in some cases, actually treating diseases involving tissue damage and destruction. One such disease, osteoarthritis, results from progressive degeneration of articular cartilage, which has a limited ability to repair itself. With tissue engineering, scientists are now able to regenerate cartilage in vitro from isolated mature chondrocytes. While the regeneration process is still not fully understood, enough has been learned that physicians are already implanting cultured chondrocytes into humans and other animals in the hopes of effecting joint repair. One aspect which has not been fully explored is the effect of mechanical stress on developing and implanted cartilage, especially over the long term. This article will review in brief what is now known about the mechanical factors affecting cartilage regeneration in vitro and what still remains to be determined for optimum tissue engineering of cartilage constructs. © 1996 John Wiley & Sons, Inc.
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  • 74
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    Biotechnology and Bioengineering 50 (1996), S. 443-451 
    ISSN: 0006-3592
    Keywords: osteoblast ; migration ; poly(αhydroxy esters) ; poly(DL-lactic-co-glycolic acid) ; PLGA ; biodegradable polymers ; tissue engineering ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We investigated the migration of rat calvaria osteoblast populations on poly(α-hydroxy ester) films for up to 14 days to determine effects of substrate composition and culture conditions on the migratory characteristics of osteoblasts. Initial osteoblast culture conditions included cell colonies formed by seeding a high (84,000 cells/cm2) or low (42,000 cells/cm2) density of isolated osteoblasts on the polymer films, and bone tissue cultures formed by plating bone chips directly on the substrates. High density osteoblast colonies cultured and allowed to migrate and proliferate radially on 85:15 poly(DL-lactic-co-glycolic acid) (PLGA) films, 75:25 PLGA films, and tissue culture polystyrene controls demonstrated that the copolymer ratio in the polymer films did not affect the rate of increase in substrate surface area (or culture area) covered by the growing cell colony. However, the rate of increase in culture area was dependent on the initial osteoblast seeding density. Initial cell colonies formed with a lower osteoblast seeding density on 75:25 PLGA resulted in a lower rate of increase in culture area, specifically 4.9 ± 0.3 mm2/day, versus 14.1 ± 0.7 mm2/day for colonies seeded with a higher density of cells on the same polymer films. The proliferation rate for osteoblasts in the high and low density seeded osteoblast colonies did not differ, whereas the proliferation rate for the osteoblasts arising from the bone chips was lower than either of these isolated cell colonies. Confocal and light microscopy revealed that the osteoblast migration occurred as a monolayer of individual osteoblasts and not a calcified tissue front. These results demonstrated that cell seeding conditions strongly affect the rates of osteoblast migration and proliferation on biodegradable poly(α-hydroxy esters). © 1996 John Wiley & Sons, Inc.
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  • 75
    ISSN: 0006-3592
    Keywords: bone marrow ; hematopoiesis ; perfusion ; culture optimization ; stroma ; stem cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hematopoiesis, the formation of mature blood cells from stem (LTC-IC) and progenitor (CFU-GM) cells in the bone marrow, is a complex tissue-forming process that leads to many important physiological functionalities. Consequently, a functioning ex vivo hematopoietic system has a variety of basic scientific and clinical uses. The design and operation of such a system presents the tissue engineer with challenges and choices. In this study, three culture variables were used to control ex vivo human hematopoiesis. Systematic variation of inoculum density (ID), medium exchange interval (MEI), and the use of preformed stroma (PFS) showed that (1) all three variables significantly influenced culture performance, (2) the three variables interacted strongly, and (3) the variables could be manipulated to achieve the optimization of different performance criteria. Donor-to-donor variability in culture performance was great at low ID but was minimized at higher ID. PFS had a large positive effect on cell and CFU-GM output at low ID, but had minimal effect at higher ID. In fact, PFS caused a decrease in LTC-IC output at high ID. The effects of PFS indicated that stromal cell elements became more limiting than proliferative cell elements as ID was reduced.In cultures without PFS, maximum cell output was obtained with high ID using a short MEI, whereas the greatest cell expansion ratio was obtained at low ID with an intermediate MEI. Maximum CFU-GM output was obtained from cultures with high ID using a short to intermediate MEI, whereas the greatest CFU-GM expansion ratio was obtained at intermediate ID with an intermediate MEI. The addition of PFS altered the locations of these maxima. In general, PFS moved the maxima to lower ID, and culture output became more sensitive to MEI. Therefore, the optimization of one performance criterion always resulted in a decline of the others. This study demonstrates that ex vivo tissue function is sensitive to many culture variables in an interactive fashion and that systematic multivariable studies are required to characterize tissue function. Once the effects of individual variables and their interactions are known, this knowledge can be used to optimize tissue performance with respect to desired criteria. © 1996 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 50 (1996) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 77
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    Biotechnology and Bioengineering 51 (1996), S. 410-421 
    ISSN: 0006-3592
    Keywords: lysozyme ; thermal stability ; 1H NMR ; conformational flexibility ; melting temperature ; PEG ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The reversible folding destabilization of hen lysozyme has been confirmed by a melting temperature (Tm) decrease in aqueous poly(ethylene glycol) (PEG). The percent denatured, extracted from the histidine 15 C2H (H15 C2H) native and denatured peak areas from 500-MHz one-dimensional proton nuclear magnetic resonance (1D 1H NMR) spectra in D2O, was analyzed through denaturation temperatures at 0% and 20% (w/w) PEG 1000. The lysozyme (3.5 mM) Tm decreased by 4.2°C and 7.1°C in 20% (w/w) PEG 1000 at pH 3.8 and 3.0, respectively. The Tm decreased with increasing lysozyme concentration. Additionally, the temperature-induced resonance migrations of 17 protons from 8 residues indicate that the native lysozyme structure undergoes temperature-induced conformational changes. The changes were essentially identical in both 0% and 20% (w/w) PEG 1000 at both pH 3.0 and 3.8. This small, local restructuring of the hydrophobic box region may be a manifestation of temperature-dependent solution hydrophobicity, whereas active-site cleft fluctuations may be due to the inherent active-site flexibility. The lysozyme structure in PEG at 35°C was determined to be essentially native from the 1H nuclear Overhauser effect spectroscopy (NOESY) fingerprint regions. Additionally, lysozyme chemical shifts, from 1D spectra, in PEG 200, 300, and 1000 at 35°C and various concentrations were essentially identical, further confirming that the conformation remains native in various PEG solutions. © 1996 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 51 (1996), S. 375-383 
    ISSN: 0006-3592
    Keywords: cellulase ; enzyme recycling ; enzyme adsorption ; lignocellulosic hydrolysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Past technoeconomic modeling work has identified the relatively large contribution that enzymatic hydrolysis adds to the total cost of producing ethanol from lignocellulosic substrates. This cost was primarily due to the high concentration of enzyme and long incubation time that was required to obtain complete hydrolysis. Although enzyme and substrate concentration and end-product inhibition influenced the rate of hydrolysis, the effect was less pronounced during the initial stages of hydrolysis. During this time most of the cellulases were adsorbed onto the unhydrolyzed residue. By recycling the cellulases adsorbed to the residual substrate remaining after an initial 24 h, a high rate of hydrolysis, with low overall residence time and minimal cellulase input, could be achieved for several rounds of enzyme recycle. A comparison of the front end (pretreatment, fractionation, and hydrolysis) of a softwood/hardwood to ethanol process indicated that the lignin associated with the softwood-derived cellulose stream limited the number of times the cellulose containing residue could be recycled. © 1996 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 51 (1996), S. 399-409 
    ISSN: 0006-3592
    Keywords: cell damage ; cell culture ; bubble aeration ; agitation ; bubble coalescence and breakup ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: It has been established that the forces resulting from bubbles rupturing at the free air (gas)/liquid surface injure animal cells in agitated and/or sparged bioreactors. Although it has been suggested that bubble coalescence and breakup within agitated and sparged bioreactors (i.e., away from the free liquid surface) can be a source of cell injury as well, the evidence has been indirect. We have carried out experiments to examine this issue. The free air/liquid surface in a sparged and agitated bioractor was eliminated by completely filling the 2-L reactor and allowing sparged bubbles to escape through an outlet tube. Two identical bioreactors were run in parallel to make comparisons between cultures that were oxygenated via direct air sparging and the control culture in which silicone tubing was used for bubble-free oxygenation. Thus, cell damage from cell-to-bubble interactions due to processes (bubble coalescence and breakup) occurring in the bulk liquid could be isolated by eliminating damage due to bubbles rupturing at the free air/liquid surface of the bioreactor. We found that Chinese hamster ovary (CHO) cells grown in medium that does not contain shear-protecting additives can be agitated at rates up to 600 rpm without being damaged extensively by cell-to bubble interactions in the bulk of the bioreactor. We verified this using both batch and high-density perfusion cultures. We tested two impeller designs (pitched blade and Rushton) and found them not to affect cell damage under similar operational conditions. Sparger location (above vs. below the impeller) had no effect on cell damage at higher agitation rates but may affect the injury process at lower agitation intensities (here, below 250 rpm). In the absence of a headspace, we found less cell damage at higher agitation intensities (400 and 600 rpm), and we suggest that this nonintuitive finding derives from the important effect of bubble size and foam stability on the cell damage process. © 1996 John Wiley & Sons, Inc.
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  • 80
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    Biotechnology and Bioengineering 51 (1996), S. 434-438 
    ISSN: 0006-3592
    Keywords: polyphosphate ; Escherichia coli ; phosphate starvation ; gene expression ; heterologous ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of intracellular polyphosphate on the phosphate-starvation response in Escherichia coli was studied by genetically manipulating the intracellular polyphosphate levels and by performing phosphate shifts on the genetically engineered strains. Strains that produced large quantities of polyphosphate and were able to degrade it induced the phosphate-starvation response to a lesser extent than wild-type strains, whereas strains that were unable to degrade a large intracellular polyphosphate pool induced the phosphate-starvation response to a greater extent than wild-type strains. These results have important implications for expression of heterologous genes under control of the phoA promoter. © 1996 John Wiley & Sons, Inc.
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  • 81
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    Biotechnology and Bioengineering 51 (1996), S. 458-465 
    ISSN: 0006-3592
    Keywords: concentric-cylinder shear device ; rotor/stator homogenization ; shear ; shear rate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Shear is present in almost all bioprocesses and high shear is associated with processes involving agitation and emulsification. The purpose of this study is to investigate the effect of high shear and high shear rate on proteins. Two concentric cylinder-based shear systems were used. One was a closed concentric-cylinder shear device (CCSD) and the other was a homogenizer with a rotor/stator assembly. Mathematical modeling of these systems allowed calculation of the shear rate and shear. The CCSD generated low shear rates (a few hundred s-1), whereas the homogenizer could generate very high shear rates (〉 105 s-1). High shear could be achieved in both systems by increasing the processing time. Recombinant human growth hormone (rhGH) and recombinant human deoxyribonuclease (rhDNase) were used as the model proteins in this study. It was found that neither high shear nor high shear rate had a significant effect on protein aggregation. However, a lower melting temperature and enthalpy were detected for highly sheared rhGH by using scanning microcalorimetry, presumably due to some changes in protein's conformation. Also, SDS-PAGE indicated the presence of low molecular-weight fragments, suggesting that peptide bond breakage occurred due to high shear. rhDNase was relatively more stable than rhGH under high shear. No conformational changes and protein fragments were observed. © 1996 John Wiley & Sons, Inc.
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  • 82
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    Biotechnology and Bioengineering 51 (1996), S. 494-499 
    ISSN: 0006-3592
    Keywords: cell metabolism ; baculovirus ; insect cells ; recombinant protein OSF-2 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The properties of Sf9 and Tn5 insect cells were analyzed comparatively under serum-free culture conditions. Sf9 cells in SF900II medium apparently utilized sucrose as a primary nutrient both before and after virus infection, yielding small amounts of lactate and ammonia. Tn5 cells in Excell 401 medium consumed all the nutrients examined, including sucrose. The productivity of a recombinant glycoprotein, OSF-2, by Tn5 cells, was moderate in both monolayer and spinner cultures, but the ability to secrete it was compromised in the former case. Relative to the Tn5 cultures, Sf9 produced 30-fold more OSF-2 in either culture mode. © 1996 John Wiley & Sons, Inc.
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  • 83
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    Biotechnology and Bioengineering 51 (1996), S. 538-543 
    ISSN: 0006-3592
    Keywords: NMR imaging ; biosorption ; alginate ; shrinking core model ; Laminaria ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this contribution, an NMR imaging study of heavy metal absorption in alginate, immobilized-cell biosorbents, and kombu (Laminaria japonica) algal biomass is presented. This method provides the good possibility of directly monitoring the time evolution of the spatial distribution of the ions in the materials. From these results, we demonstrate that rare earth ions are absorbed with a steep reaction front that can be described very well with a modified shrinking core model, while copper ions are absorbed with a more diffuse front.
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  • 84
    ISSN: 0006-3592
    Keywords: oxidoreductase ; chiral alcohol ; racemic resolution ; membrane reactor ; continuous extraction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Oxidations of alcohols by alcohol dehydrogenases often suffer from low conversions and slow reaction rates due to severe product inhibition. This can be overcome by continuous product extraction, because only the concentrations, but not the kinetic parameters, can be changed. As a consequence, it is favorable to apply a differential circulation reactor with continuous product extraction, where only a small amount of product is formed per cycle. The product is then directly extracted using a microporous hydrophobic hollow fiber membrane. This results in an increase of the relative activity of the dehydrogenase at a given conversion. The reaction investigated is the kinetic resolution of racemic 1-phenyl-1,2-ethanediol by glycerol dehydrogenase (GDH). The resulting oxidation product, 2-hydroxyacetophenone, causes a strong product inhibition. Additionally, it reacts in a chemical reaction with the cofactor lowering its active concentration. Because the GDH needs β-nicotinamide adenine dinucleotide (NAD+) as a cofactor, lactate dehydrogenase is used to regenerate NAD+ from NADH by reducing pyruvate to (L)-lactate. A conversion of 50% with respect to the racemate and an enantiomeric excess 〉99% of the (S)-enantiomer was reached.
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  • 85
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    Biotechnology and Bioengineering 51 (1996), S. 581-590 
    ISSN: 0006-3592
    Keywords: microfiber ; graft polymerization ; DNA immobilization ; immunoadsorbent ; DNA ; anti-DNA antibody ; systemic lupus erythematosus ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Immobilization of DNA to the surface of poly(ethylene terephthalate) (PET) microfibers with a high specific surface area of 0.83 m2/g was carried out to give the fiber surface an affinity for anti-DNA antibody. Following ozone oxidation, the microfibers were subjected to graft polymerization of monomers including acrylic acid, methacryloyloxyethyl phosphate, N,N-dimethylaminoethyl methacrylate, N-vinylformamide, and glycidyl methacrylate. Calf thymus DNA was immobilized to the grafted fiber surface through either covalent binding or polyion complexation with the grafted polymer chains. The highest surface density of DNA immobilized (0.6 μg/cm2) was obtained when DNA was immobilized through formation of phosphodiester linkage between the hydroxyl group of DNA and the phosphate group in grafted poly(methacryloyloxyethyl phosphate) using 1,1-carbonyldiimidazole, or through polyion complexation between the anionic DNA and the cationic grafted poly(N,N-dimethylaminoethyl methacrylate) chains. Batch adsorption of anti-DNA antibody to the grafted PET fibers with and without DNA immobilized on their surface was conducted with serum obtained from systemic lupus erythematosus model mice. The DNA-immobilized PET fibers exhibited a higher adsorption capacity and specificity than the others. In addition, the DNA-immobilized fibers effectively adsorbed human anti-DNA antibody.
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  • 86
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    Biotechnology and Bioengineering 49 (1996), S. 20-25 
    ISSN: 0006-3592
    Keywords: filamentous fungi ; immobilization ; biofilm bioreactor ; oil emulsion ; degradation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new type of horizontal biofilm bioreactor for continuous bioconversion of emulsified oily substrate by immobilized growing biofilm of filamentous fungi was designed, constructed, and feasibility tested. The new reactor design provides “self”-immobilization of homogenized mycelium leading to even biofilm development. This was accomplished by using stainless steel screens of optimal mesh, mounted in parallel and stretching outward from a main rotating axis of a biological rotating contractor. Each screen was equipped with a pair of stainless steel blades mounted on supports allowing for continuous biofilm “shaving” beyond a predetermined thickness, thus retaining freshly growing active biofilm surface. The feasibility of the new bioreactor was demonstrated by decalactone production from emulsified castor oil by immobilized filamentous fungi (Tyromyces sambuceus). The combination of oriented metal screens and moving blades was found to be highly effective for a model system in maintaining stable substrate emulsion in the reactor in either batchwise or continuous processing, as well as maintaining biofilm thickness with continuous removal of excess growing hyphae. © 1996 John Wiley & Sons, Inc.
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  • 87
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    Biotechnology and Bioengineering 49 (1996), S. 284-289 
    ISSN: 0006-3592
    Keywords: organic phase biocatalysis ; esterification ; water activity ; lipase ; biocatalysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Packed bed hollow fiber membrane reactors were used to carry out organic phase biocatalysis at constant water activity. The performance of the device was tested by carrying out the esterification of dodecanol and decanoic acid in hexane. Lipase from Candida rugosa, immobilized on microporous polypropylene and packed in the shell space of the reactor, was used to catalyze the reaction. In situ water activity control was accomplished by pumping appropriate saturated salt solutions through the microporous hollow fiber polypropylene membranes. Water generated by reaction in the organic phase, pumped continuously through the shell of the reactor, was transferred into the bulk of the aqueous phase under the water activity gradient. The reactor performance was found to be strongly dependent on the controlling water activity. By carefully selecting this control activity it was found possible to obtain complete esterification. The water activity of the organic phase could be maintained very close to that of the saturated salt solution used. The reactor could be operated in the continuous mode for 100 h without any degradation in its performance. © 1996 John Wiley & Sons, Inc.
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  • 88
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    Biotechnology and Bioengineering 49 (1996), S. 328-333 
    ISSN: 0006-3592
    Keywords: disaccharides ; lipase ; organic solvent ; trans-esterification ; regioselectivity ; fatty acid ester ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The acylation of several disaccharides by ethyl butanoate and ethyl dodecanoate was catalyzed by Candida antarctica lipase in tert-butyl alcohol, at temperatures ranging from 40° to 82°C (reflux temperature). The relative reaction rates of the various disaccharides were directly related to their solubility. The primary products were the monoesters derived from acylation of the primary alcohol groups. At higher conversions diesters were formed, and the ratio of diester to monoester was markedly dependent on the structure of the disaccharide. Thus, reaction of maltose with ethyl dodecanoate in refluxing tert-butyl alcohol afforded the 6′-monododecanoate even at high conversions. Trehalose, in contrast, afforded the 6,6′-diester. Acylation of the less soluble sucrose and lactose was much slower, but a moderate (37%) conversion of sucrose was observed after a prolonged reaction time (7 days). A number of other lipases and proteases were tested but C. antarctica lipase was unique in catalyzing the acylation of sucrose in refluxing tert-butyl alcohol. © 1996 John Wiley & Sons, Inc.
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  • 89
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    Biotechnology and Bioengineering 49 (1996), S. 341-347 
    ISSN: 0006-3592
    Keywords: water reuse ; fermentation ; lysine ; Corynebacterium glutamicum ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: L-Lysine is produced commercially by fermentation. As is typical for fermentation processes, a large amount of liquid waste is generated. To minimize the waste, which is mostly the broth effluent from the cation exchange column used for l-lysine recovery, we investigated a strategy of recycling a large fraction of this broth effluent to the subsequent fermentation. This was done on a labscale process with Corynebacterium glutamicum ATCC 21253 as the l-lysine-producing organism. Broth effluent from a fermentation in a defined medium was able to replace 75% of the water for the subsequent batch; this recycle ratio was maintained for three sequential batches without affecting cell mass and l-lysine production. Broth effluent was recycled at 50% recycle ratio in a fermentation in a complex medium containing beet molasses. The first recycle batch had an 8% lower final l-lysine level, but 8% higher maximum cell mass. In addition to reducing the volume of liquid waste, this recycle strategy has the additional advantage of utilizing the ammonium desorbed from the ion-exchange column as a nitrogen source in the recycle fermentation. The major problem of recycling the effluent from the complex medium was in the cation-exchange operation, where column capacity was 17% lower for the recycle batch. The loss of column capacity probably results from the buildup of cations competing with l-lysine for binding. © 1996 John Wiley & Sons, Inc.
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  • 90
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    Biotechnology and Bioengineering 49 (1996), S. 377-382 
    ISSN: 0006-3592
    Keywords: hybridoma ; batch culture ; dichloroacetate ; metabolism ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We have studied the effect of the pyruvate dehydrogenase (PDH) activator, dichloroacetate (DCA), on the growth, metabolism, and productivity of the PQXB ½ hybridoma cell line. In control batch cultures, cessation of growth and the onset of decline phase coincided with the time at which the media became exhausted of glutamine. Supplementation of the media with DCA (1 mM) extended the growth phase of this cell line by approximately 20 h without affecting its growth rate. This prolonged period of growth resulted in an increased maximum cell density (16%) and final antibody yield (55%). Repeat experiments showed these effects to be reproducible, with the increases in antibody yield being between 50 and 60%. DCA did not affect the specific rates of glucose utilization and lactate production. However, it decreased the specific glutamine consumption rate. This characteristic of DCA action appeared, at least in part, to provide an explanation for the extended growth phase exhibited by DCA-treated cultures, since it delayed the time at which the media became depleted of glutamine. The consumption and production kinetics for various nutrients and their metabolites in both control and DCA-treated cultures suggested that: (1) glutamine catabolism proceeded by a pathway involving conversion to glutamate by glutaminase followed by subsequent transamination by alanine aminotransferase, and (2) DCA decreased the specific glutamine consumption rate by directly or indirectly inhibiting the transamination. It is expected that the routine inclusion of DCA in media used for hybridoma cultivation will be valuable for enhancement of monoclonal antibody (Mab) yields on a laboratory scale. © 1996 John Wiley & Sons, Inc.
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  • 91
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    Biotechnology and Bioengineering 49 (1996), S. 399-404 
    ISSN: 0006-3592
    Keywords: subtilisin ; hydration ; catalytic activity ; supercritical fluids ; solvent effects ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We studied a model transesterification reaction catalyzed by subtilisin Carlsberg suspended in carbon dioxide, propane, and mixtures of these solvents under pressure. To account for solvent effects due to differences in water partitioning between the enzyme and the bulk solvents, we measured water sorption isotherms for the enzyme in each solvent. We measured catalytic activity as a function of enzyme hydration and obtained bell-shaped curves with maxima at the same enzyme hydration (12%) in all the solvents. However, the activity maxima were different in all media, being much higher in propane than in either CO2 or the mixtures with 50 and 10% CO2. Considerations based on the solvation ability of the solvents did not offer an explanation for the differences in catalytic activity observed. Our results suggest that CO2 has a direct adverse effect on the catalytic activity of subtilisin. © 1996 John Wiley & Sons, Inc.
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  • 92
    ISSN: 0006-3592
    Keywords: Streptomyces virginiae ; autoregulator ; virginiae butanolide ; virginiamycin fermentation ; optimization ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A strategy for optimization of non-growth-associated production in batch culture employing an empirical approach was developed through the study of virginiamycin production. The strategy is formulated with two aims: attaining a high cell concentration at the beginning of the production phase without decrease in production activity; and enhancing the production activity during the production phase. As a practical example, the goal of a maximum virginiamycin (M and S) production in the batch culture of Streptomyces virginiae was set. To attain a high cell concentration in the production phase of the batch culture, that is, to extend the growth phase for as long as possible, the optimum composition and concentration of the complex medium, especially the yeast extract (YE) concentration, were first investigated. Dissolved oxygen (DO) concentration control was also a parameter considered in maintaining the production activity during the production phase. In addition, to enhance the production activity, an optimum addition strategy of an autoregulator, virginiae butanolide-C (VB-C), was investigated. Combining these measures, the optimum cultivation conditions were found to be an initial YE concentration in the complex medium of 45 g/L, the shot addition of 300 μg/L of VB-C 11.5 h after the start of the batch culture, and a DO concentration maintained above 2 mg/L. The maximum concentrations of virginiamycin M and S were about ninefold those obtained under nonoptimum cultivation conditions. Nonoptimum cultivation conditions consisted of an initial YE concentration one sixth (7.5 g/L) that of the optimum cultivation conditions, and no VB-C addition. These conditions were used as representative of the standard cultivation of virginiamycin in this study. The strategy developed here will be applicable to the production of other antibiotics, especially to the cultivation of Streptomyces species, in which a hormonelike signal material (an autoregulator) plays an important role in antibiotic production. © 1996 John Wiley & Sons, Inc.
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  • 93
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    Biotechnology and Bioengineering 49 (1996) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 94
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    Biotechnology and Bioengineering 49 (1996), S. 504-511 
    ISSN: 0006-3592
    Keywords: cellulose ; cellulase ; simultaneous saccharification and extractive fermentation ; ethanol ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Alcohol fermentation has traditionally been carried out in aqueous environments because of the ready solubility of reactant (sugar) and product (ethanol). However, extraction of the product ethanol into a nonmiscible phase can result in kinetic benefits due to reduced inhibition of the fermentation reactions. In this study, we report the development of a novel simultaneous saccharification and extractive fermentation (SSEF) process. Ethanol productivity was increased by up to 65% over conventional (nonextractive) fed-batch simultaneous saccharification systems when calculated on the basis of aqueous phase volume. The amount of water required for SSEF reactions was dramatically reduced from that required for conventional SSF. In batch SSEF reactors with 2.5% aqueous phase, 50% conversion of 25% (aqueous phase concentration) Solka Floc could be achieved in 48 h using 2 FPU/g cellulase. © 1996 John Wiley & Sons, Inc.
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  • 95
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    Biotechnology and Bioengineering 49 (1996), S. 535-543 
    ISSN: 0006-3592
    Keywords: whole cell biotransformation ; biocatalyst ; baker's yeast ; immobilization ; microencapsulation ; organic solvents ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Stable, semipermeable polyamide microcapsules were prepared by interfacial polymerization from a mixture of 1,6-hexanediamine and poly(allylamine) crosslinked with di-acid chlorides and were used to encapsulate baker's yeast. The size and distribution of cells within the capsules were investigated by a combination of laser confocal, electron scanning, and transmission electron microscopy. The encapsulated cells were studied as a biocatalyst for the model reduction of 1-phenyl-1,2-propanedione to 2-hydroxy-1-phenyl-1-propanone in a number of organic solvents. The polymerization conditions were extensively investigated and were found to greatly influence the product yield. Microencapsulated yeast cells, prepared under optimized conditions, carried out the reduction more efficiently than free cells as well as those immobilized in alginate and κ-carrageenan beads. The developed methodology should be broadly applicable to other biotransformations of interest. © 1996 John Wiley & Sons, Inc.
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  • 96
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    Biotechnology and Bioengineering 49 (1996), S. 587-598 
    ISSN: 0006-3592
    Keywords: gas phase bioreactor ; dynamic behavior ; waste air ; MEK ; MIBK ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In the work reported here, selected aspects of the dynamic behavior of biofilters for waste air treatment have been investigated. Emphasis was placed on transient state elimination of mixtures of methyl ethyl ketone (MEK) and methyl isobutyl ketone (MIBK) vapors and on explanation of the observed phenomena. The initial startup, the response of the biofilter to step changes in the pollutant loadings, responses to pollutant pulses, restarting after starvation, and the influence of step changes in gaseous phase oxygen partial pressure are presented and discussed. © 1996 John Wiley & Sons, Inc.
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    Biotechnology and Bioengineering 49 (1996), S. 621-628 
    ISSN: 0006-3592
    Keywords: Kluyveromyces ; Candida utilis ; Kluyver effect ; chemostat ; biomass ; whey ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Many facultatively fermentative yeast species exhibit a “Kluyver effect”: even under oxygen-limited growth conditions, certain disaccharides that support aerobic, respiratory growth are not fermented, even though the component monosaccharides are good fermentation substrates. This article investigates the applicability of this phenomenon for high-cell-density cultivation of yeasts. In glucose-grown batch cultures of Candida utilis CBS 621, the onset of oxygen limitation led to alcoholic fermentation and, consequently, a decrease of the biomass yield on sugar. In maltose-grown cultures, alcoholic fermentation did not occur and oxygen-limited growth resulted in high biomass concentrations (90 g dry weight L-1 from 200 g L-1 maltose monohydrate in a simple batch fermentation). It was subsequently investigated whether this principle could also be applied to Kluyveromyces species exhibiting a Kluyver effect for lactose. In oxygen-limited, glucose-grown chemostat cultures of K. wickerhamii CBS 2745, high ethanol concentrations and low biomass yields were observed. Conversely, ethanol was absent and biomass yields on sugar were high in oxygen-limited chemostat cultures grown on lactose. Batch cultures of K. wickerhamii grown on lactose exhibited the same growth characteristics as the maltose-grown C. utilis cultures: absence of ethanol formation and high biomass yields. Within the species K. marxianus, the occurrence of a Kluyver effect for lactose is known to be strain dependent. Thus, K. marxianus CBS 7894 could be grown to high biomass densities in lactose-grown batch cultures, whereas strain CBS 5795 produced ethanol after the onset of oxygen limitation and, consequently, yielded low amounts of biomass. Because the use of yeast strains exhibiting a Kluyver effect obviates the need for controlled substrate-feeding strategies to avoid oxygen limitation, such strains should be excellently suited for the production of biomass and growth-related products from low-cost disaccharide-containing feedstocks. © 1996 John Wiley & Sons, Inc.
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  • 98
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    Biotechnology and Bioengineering 49 (1996), S. 659-666 
    ISSN: 0006-3592
    Keywords: suspension culture ; insect cells ; baculovirus ; multiplicity of infection ; time of infection ; model ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In vitro infection of insect cells with baculoviruses is increasingly considered a viable means for the production of biopesticides, recombinant veterinary vaccines, and other recombinant products. Batch fermentation processes traditionally employ intermediate to high multiplicities of infection necessitating two parallel scale-up processes - one for cells and one for virus. In this study, we consider the use of multiplicities of infection as low as 0.0001 plaque-forming units per cell, a virus level low enough to enable infection of even large reactors (e.g., 10 m3) directly from a frozen stock. Using low multiplicities in the Sf9/β-gal-AcNPV system, recombinant protein titers comparable with the maximum titer observed in high multiplicity infections were achieved. Cultures yielding the maximum titer were characterized by reaching a maximum cell density between 3 and 4 × 109 cell L-1. This optimal cell yield did not depend on the multiplicity of infection, supporting the existing view that batch cultures are limited by availability of substrate. Up to a certain cell density, product titer will increase almost linearly with availability of biocatalyst, that is, cells. Beyond this point any further cell formation comes at the expense of final product titer. Low multiplicity infections were found not to cause any significant dispersion of the protein production process. Hence, product stability is not a major issue of concern using low multiplicities of infection. The sensitivity to initial conditions and disturbances, however, remains an issue of concern for the commercial use of low multiplicity infections. © 1996 John Wiley & Sons, Inc.
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  • 99
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    Biotechnology and Bioengineering 49 (1996), S. 700-708 
    ISSN: 0006-3592
    Keywords: alcohol dehydrogenase ; enzyme ; water sorption isoterm ; gas phase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The adsorption of water by alcohol dehydrogenase from baker's yeast (YADH) has been measured in a continuous-flow gas reactor at varying temperatures. Adsorption isotherms in the presence of gaseous organic substrates are compared to those from organic-free gas mixtures. Almost no effect of the hydrophobic molecule on total water adsorption was observed. A rarely mentioned multilayer isotherm model from the 1930s, the Huttig's isotherm, has been found to fit the experimental data with extremely good accuracy. The model enables the calculation of both the heat of adsorption of water to the enzyme and the total amount of water necessary for monolayer coverage. The heat of adsorption of water in the first layer is approximately -16 kcal/mol. This tight binding of water, which is much higher than the heat of condensation of pure water, helps to explain the kinetic properties of YADH-catalyzed reactions on vapor phase substrates. While the monolayer coverage is temperature independent, the enzyme demonstrates hysteresis when transitioning between adsorption and desorption. The hysteresis observed in water sorption studies may also explain previously reported properties of the enzyme. © 1996 John Wiley & Sons, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 699-699 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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