Recovery of Poly-β-Hydroxybutyrate from Estuarine Microflora

Poly-β-hydroxybutyrate (PHB) is a uniquely procaryotic endogenous storage polymer whose metabolism has been shown to reflect environmental perturbations in laboratory monocultures. When hydrolyzed for 45 min in 5% sodium hypochlorite, PHB can be isolated from estuarine detrital microflora in high yield and purified free from non-PHB microbial components. Lyophilization of frozen estuarine samples shortens the exposure time to NaOCl necessary for maximal recovery. Lyophilized samples of hardwood leaves, Vallisneria, and the aerobic upper millimeter of estuarine muds yielded PHB. The efficiency of incorporation of sodium [1-14C]acetate into PHB is very high and is stimulated by aeration. PHB was not recovered from the anaerobic portions of sediments unless they were aerated for a short time. Levels of PHB in the detrital microbial community do not correlate with the microbial biomass as measured by the extractible lipid phosphate, suggesting that PHB-like eucaryotic endogenous storage materials may more accurately reflect the metabolic status of the population than its biomass.

[1]  G. Gottschalk,et al.  Formation and Utilization of Poly-β-Hydroxybutyric Acid by Knallgas Bacteria (Hydrogenomonas) , 1961, Nature.

[2]  A. C. Emeruwa,et al.  Poly-β-Hydroxybutyrate Metabolism During Growth and Sporulation of Clostridium botulinum , 1973, Journal of bacteriology.

[3]  J. Davis.  CELLULAR LIPIDS OF A NOCARDIA GROWN ON PROPANE AND N-BUTANE. , 1964, Applied microbiology.

[4]  W. J. Dyer,et al.  A rapid method of total lipid extraction and purification. , 1959, Canadian journal of biochemistry and physiology.

[5]  W. Wiessner,et al.  Minimum Requirements for Heterotrophic Growth, and Reserve Substance in Beggiatoa , 1963, Nature.

[6]  G. Rohlich,et al.  A mechanism of floc formation by Zoogloea ramigera. , 1966, Journal - Water Pollution Control Federation.

[7]  CHARACTERIZATION OF POLY-BETA-HYDROXYBUTYRATE EXTRACTED FROM DIFFERENT BACTERIA. , 1965, Journal of bacteriology.

[8]  S. J. Morrison,et al.  Biochemical Measurements of Microbial Mass and Activity from Environmental Samples , 1979 .

[9]  Kenneth L. Burdon,et al.  Fatty Material in Bacteria and Fungi Revealed by Staining Dried, Fixed Slide Preparations , 1946, Journal of bacteriology.

[10]  D. White,et al.  Muramic acid as a measure of microbial biomass in estuarine and marine samples , 1977, Applied and environmental microbiology.

[11]  N. Carr.  The occurrence of poly-beta-hydroxybutyrate in the blue-green alga, Chlorogloea fritschii. , 1966, Biochimica et biophysica acta.

[12]  S. J. Morrison,et al.  Effects of Substrate Biodegradability on the Mass and Activity of the Associated Estuarine Microbiota , 1978, Applied and environmental microbiology.

[13]  E. Dawes,et al.  A disk assay for poly- -hydroxybutyrate. , 1973, Analytical biochemistry.

[14]  J. Wilkinson,et al.  Poly-beta-hyroxybutyrate metabolism in washed suspensions of Bacillus cereus and Bacillus megaterium. , 1958, Journal of general microbiology.

[15]  J. Sobek,et al.  Endogenous Metabolism of Azotobacter agilis , 1966, Journal of bacteriology.

[16]  S. J. Morrison,et al.  Determination of microbial activity of estuarine detritus by relative rates of lipid biosynthesis1 , 1977 .

[17]  D. Williamson,et al.  The isolation and estimation of the poly-beta-hydroxybutyrate inclusions of Bacillus species. , 1958, Journal of general microbiology.

[18]  G. Rohlich,et al.  Isolation, Identification, and Metabolic Role of the Sudanophilic Granules of Zoogloea ramigera , 1965, Applied microbiology.

[19]  M. D. Socolofsky,et al.  Cyst Formation and Poly-β-Hydroxybutyric Acid Accumulation in Azotobacter , 1966 .