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  • 1
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] The alk genes from the catabolic OCT plasmid of Pseudomonas oleovorans, which encode the enzymes involved in the oxidation of n-alkanes to carboxylic acids, were introduced into E. coli W3110. The resulting recombinant converts n-octane in a two-liquid phase medium into the corresponding alkanoate ...
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  • 2
    ISSN: 0006-3592
    Keywords: Pseudomonas oleovorans ; poly(3-hydroxyalkanoate) ; n-octane ; high cell density ; fed-batch culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pseudomonas oleovorans is able to accumulate poly(3-hydroxyalkanoates) (PHAs) under conditions of excess n-alkanes, which serve as sole energy and carbon source, and limitation of an essential nutrient such as ammonium. In this study we aimed at an efficient production of these PHAs by growing P. oleovorans to high cell densities in fed-batch cultures.To examine the efficiency of our reactor system, P. oleovorans was first grown in batch cultures using n-octane as growth substrate and ammonia water for pH regulation to prevent ammonium limiting conditions. When cell growth ceased due to oxygen limiting conditions, a maximum cell density of 27 g ·L-1 dry weight was obtained. When the growth temperature was decreased from the optimal temperature of 30°-18°C, cell growth continued to a final cell density of 35 g · L-1 due to a lower oxygen demand of the cells at this lower incubation temperature.To quantify mass transfer rates in our reactor system, the volumetric oxygen transfer coefficient (kLa) was determined during growth of P. oleovorans on n-octane. Since the stirrer speed and airflow were increased during growth of the organism, the kLa also increased, reaching a constant value of 0.49 s-1 at maximum airflow and stirrer speed of 2 L · min-1 and 2500 rpm, respectively. This kLa value suggests that oxygen transfer is very efficient in our stirred tank reactor.Using these conditions of high oxygen transfer rates, PHA production by P. oleovorans in fed-batch cultures was studied. The cells were first grown batchwise to a density of 6 g · L-1, after which a nutrient feed, consisting of (NH4)2SO4 and MgSO4, was started. The limiting nutrient ammonium was added at a constant rate of 0.23 g NH4+ per hour, and when after 38 h the feed was stopped, a biomass concentration of 37.1 g · L-1 was obtained. The Cellular PHA content was 33% (w/w), which is equal to a final PHA yield of 12.1 g · L-1 and an overall PHA productivity of 0.25 g PHA produced per liter medium per hour. © 1993 John Wiley & Sons, Inc.
    Additional Material: 4 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 52 (1996), S. 301-308 
    ISSN: 0006-3592
    Keywords: two-liquid phase biocatalysis ; alkane hydroxylase ; xylene oxygenase ; octanoic acid ; styrene oxide ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The Pseudomonas oleovorans alkane hydroxylase and xylene oxygenase from Pseudomonas putida are versatile mono-oxygenases for stereo- and regioselective oxidation of aliphatic and aromatic hydrocarbons. Pseudomonas oleovorans and alkanol dehydrogenase deficient mutants of Pseudomonas have previously been used to produce alkanols from various alkanes and optically active epoxides from alkenes. Similarly, P. putida strains have been used to produce aromatic alcohols, aromatic acids, and optically active styrene oxides. A limitation in the use of Pseudomonas strains for bioconversions is that these strains can degrade some of the products formed. To counter this problem, we have constructed Escherichia coli recombinants, which contain the alk genes from the OCT plasmid of P. oleovorans [E. coli HB101 (pGEc47)] and the xylMA genes from the TOL plasmid of P. putida mt-2 [E. coli HB101 (pGB63)], encoding alkane hydroxylase and xylene oxygenase, respectively. Escherichia coli HB101 (pGEc47) was used to produce octanoic acid from n-octane and E. coli HB101 (pBG63) was put to use for the oxidation of styrene to styrene oxide in two-liquid phase biocatalysis at high cell densities. The alk+ recombinant strain E. coli HB101 (pGEc47) was grown to 40 g/L cell dry mass in the presence of n-octane, which was converted to octanoic acid by the alkane oxidation system, the product accumulating in the aqueous phase. The xyl+ recombinant E. coli HB101 (pBG63) was grown to a cell density of 26 g/L cell dry mass in the presence of around 7% (v/v) n-dodecane, which contained 2% (v/v) styrene. The recombinant E. coli (xyl+) converted styrene to (S)-(+)-styrene oxide at high enantiomeric excess (94% ee) and this compound partitioned almost exclusively into the organic phase. Using these high-cell-density two-liquid-phase cultures, the products accumulated rapidly, yielding high concentrations of products (50 mM octanoic acid and 90 mM styrene oxide) in the respective phases. © 1996 John Wiley & Sons, Inc.
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  • 4
    Publication Date: 1991-04-01
    Print ISSN: 0733-222X
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Published by Springer Nature
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  • 5
    Publication Date: 1992-11-01
    Print ISSN: 0141-0229
    Electronic ISSN: 1879-0909
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Published by Elsevier
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