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Suspension cultivation of Mycobacterium ulcerans for the production of mycolactones (2001)

Cadapan, L.D. ; Arslanian, R.L. ; Carney, J.R. ; [et al.]

Oxford, UK : Blackwell Publishing Ltd

FEMS microbiology letters 205 (2001), S. 0

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ISSN: 1574-6968

Source: Blackwell Publishing Journal Backfiles 1879-2005

Topics: Biology

Notes: Mycolactones are polyketide toxins produced by Mycobacterium ulcerans, the causative agent of the tropical skin disease known as Buruli ulcer. Development of novel therapeutic agents from mycolactones has been hindered by the difficulty of producing sufficient amounts of material. Here, we describe the successful adaptation of M. ulcerans to suspension cultivation and the development of a fed-batch fermentation process that was scaled up to 150 l. In addition to producing mycolactones A and B, a number of new mycolactone-related compounds were also observed.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1111/j.1574-6968.2001.tb10977.x

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Modeling the population dynamics of baculovirus-infected insect cells: Optimizing infection strategies for enhanced recombinant protein yields (1992)

Licari, P. ; Bailey, J. E.

New York, NY [u.a.] : Wiley-Blackwell

Biotechnology and Bioengineering 39 (1992), S. 432-441

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ISSN: 0006-3592

Keywords: nuclear polyhedrosis virus ; polyhedrin promoter ; population dynamics ; multiplicity of infection ; Poisson distribution ; baculovirus ; Chemistry ; Biochemistry and Biotechnology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology

Notes: The insect cell-baculovirus model presented here is capable of simulating cell population dynamics, extracellular virion densities, and heterologous product titers in reasonable agreement with experimental data for a wide rang of multiplicities of infection (MOI) and times of infection. The model accounts for the infection of a single cell by multiple virions and the consequences on the time course of infection. The probability of infection by more than one virion was approximated using the Poisson distribution, which proved to be a refinement over second-order kinetics. The model tracks initiation and duration of important events in the progression of infected cell development (virus replication, recombinant protein synthesis, and cell lysis) for subpopulations delineated by the time and extent of their initial infection. The model suggests infection strategies, weighing the importance of MOI and infection time. Maximum product titers result from infection in the early exponential growth phase with low MOI.

Additional Material: 7 Ill.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1002/bit.260390409

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Factors influencing recombinant protein yields in an insect cell-bacuiovirus expression system: Multiplicity of infection and intracellular protein degradation (1991)

Licari, P. ; Bailey, J. E.

New York, NY [u.a.] : Wiley-Blackwell

Biotechnology and Bioengineering 37 (1991), S. 238-246

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ISSN: 0006-3592

Keywords: Chemistry ; Biochemistry and Biotechnology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology

Notes: The insect cell (Sf9)-baculovirus (AcNPV) expression system was employed for the synthesis of β-galactosidase, a model heterologous protein. In the recombinant virus studied, the lacZ gene is fused to a portion of the polyhedrin structural gene and is under the control of the polyhedrin promoter. The effect of the multiplicity of infection (MOI) on product titer was determined by infecting cells with MOI values ranging from 0 to 100 and monitoring the production of β-galactosidase with time. The relationship between final product titer and MOI was dependent on the growth phase of the cells prior to infection. The final product titer from cells infected in the early exponential phase was relatively independent of MOI. For cells infected in late-exponential phase there was a logarithmic relationship between the final β-galactosidase titer and the MOI used, with the highest MOI studied resulting in greatest protein synthesis. The synthesis and degradation rates of β-galactosidase were investigated by a pulse-chase technique using L-[35S]-methionine. At 24 h postinfection, the degradation rate is of the same order of magnitude as the synthesis rate. However, the synthesis rate of β-galactosidase increases dramatically at 96 h postinfection. During this later period, the degradation rate is negligible. Although degradation of recombinant protein occurs in this system, degradation activity declines as infection proceeds and is insignificant late in intention when recombinant protein expression is intense.

Additional Material: 10 Ill.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1002/bit.260370306

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Production of a discrete, heterogeneous population of β-galactosidase polypeptides using baculovirus expression vectors (1992)

Licari, P. ; Bailey, J. E.

New York, NY [u.a.] : Wiley-Blackwell

Biotechnology and Bioengineering 39 (1992), S. 932-944

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ISSN: 0006-3592

Keywords: Spodoptera frugiperda ; Autographa californica ; nuclear polyhedrosis virus ; polyhedrin promoter ; β-galactosidase ; heterogeneous polypeptides ; Chemistry ; Biochemistry and Biotechnology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology

Notes: Gel electrophoresis analysis of immunoprecipitated β-galactosidase and polyhedrin-β-galactosidase expressed in Spodoptera frugiperda cells infected with recombinant Autograph californica nuclear polyhedrosis virus revealed the existence of a population of discrete β-galactosidase polypeptides. Several of the polypeptides observed in the fusion protein expression experiments exhibit a consistent pattern of slightly greater molecular weight when compared to the nonfusion β-galactosidase that is compatible with the hypothesis that these fusion protein fragments retain the N-terminal polyhedrin residues. Pulse-chase experiments showed that overall β-galactosidase degradation occurred at a negligible rate compared to the synthesis rate at 96 h postinfection, yet the fragments are observed for short pulse times. Degradation of several different β-galactosidase polypeptides was observed 24 h postinfection. Ribonucleic acid hybridization analysis of lacZ transcripts shows significant heterogeneity that may result from premature transcription termination. Although a proteolytic origin cannot be excluded, the data assembled suggest that premature termination of transcription or translation is the likely cause for the heterogeneous population of immunoreactive peptides observed. Many discrete forms of β-galactosidase polypeptides were also observed in studies with Escherichia coli, indicating that production of these heterogeneous forms is not a consequence of heterologous expression of the enzyme.

Additional Material: 10 Ill.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1002/bit.260390908

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