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Appl Environ Microbiol. 1989 June; 55(6): 1334-1339
Copyright © 1989, American Society for Microbiology. All Rights Reserved.

Hyperproduction of Poly-ß-Hydroxybutyrate during Exponential Growth of Azotobacter vinelandii UWD

William J. Page^* and Olga Knosp

Department of Microbiology, University of Alberta, Edmonton, Alberta, Canada T6G 2E9

ABSTRACT

The transformation of Azotobacter vinelandii UW with A. vinelandii 113 DNA resulted in the formation of rifampin-resistant colonies, 13% of which also inherited a previously unrecognized mutation in the respiratory NADH oxidase. These transformants produced colonies with a white-sectored phenotype after prolonged incubation. Cells from these sectors were separated and purified by streaking and were named UWD. The dense white phenotype was due to the production of a large amount of poly-ß-hydroxybutyrate during the exponential growth of strain UWD. The polymer accounted for 65 or 75% of the cell dry weight after 24 h of incubation of cultures containing glucose and either ammonium acetate or N₂, respectively, as the nitrogen source. Under the same conditions, strain UW cells contained 22 to 25% poly-ß-hydroxybutyrate, but O₂-limited growth was required for these optimal production values. Polymer production was not dependent on O₂ limitation in strain UWD, but the efficiency of conversion of glucose to poly-ß-hydroxybutyrate was enhanced in O₂-limited cultures. Conversion efficiencies were >0.25 and 0.33 mg of poly-ß-hydroxybutyrate per mg of glucose consumed under vigorous- and low-aeration conditions, respectively, compared with an efficiency of 0.05 achieved by strain UW. Strain UWD, therefore, appeared to from poly-ß-hydroxybutyrate under novel conditions, which may be useful in designing new methods for the industrial production of biodegradable plastics.

^* Corresponding author.

Present address: Institut für Medizinische Chemie and Biochemie der Universität Innsbruck, A-6020 Innsbruck, Austria.

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