- Volume 19, Issue 1, 1958
Volume 19, Issue 1, 1958
- Article
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The Isolation and Estimation of the Poly-β-hydroxy-butyrate Inclusions of Bacillus Species
More LessSUMMARY: Treatment of the cells of various Bacillus spp. with an alkaline solution of sodium hypochlorite resulted in dissolution of the cells and liberation of the intracellular lipid inclusion bodies. Analyses of the isolated and purified inclusions of Bacillus cereus grown under a variety of cultural conditions showed them to contain about 89 % poly-β-hydroxybutyrate and 11% ether-soluble lipid. Parallel estimations of the poly-β -hydroxybutyrate content of intact organisms by Lemoigne’s chloroform extraction method showed that all of it was present in the lipid inclusions. These observations form the basis of a simple and rapid method of estimating the poly-β-hydroxybutyrate content of Bacillus spp. It consists essentially of digesting a washed bacterial suspension with a standard alkaline hypochlorite solution under standard conditions and measuring the residual turbidity.
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Poly-β-hyroxybutyrate Metabolism in Washed Suspensions of Bacillus cereus and Bacillus megaterium
More LessSUMMARY: Poly-β-hy droxybutyrate has been previously shown to be a major component of bacterial ‘lipid’ granules. In the present study, the conditions under which it was formed and degraded by Bacillus cereus and B. megaterium were studied in washed suspensions. Suitable substrates for synthesis were glucose, pyruvate or β-hy droxybutyrate. Acetate, although alone unable to induce synthesis, greatly enhanced formation in presence of these substrates. Under optimal conditions, suspensions synthesized up to eight times their original content of poly-β-hydroxy- butyrate in 4 hr. Formation was inhibited by high concentrations of oxygen, although no synthesis occurred anerobically in nitrogen. The optimal concentration of oxygen was about 5% . B. cereus only was able to synthesize poly-β-hydroxybutyrate in an atmosphere of hydrogen. In the absence of an external carbon and energy source, degradation occurred rapidly aerobically, to carbon dioxide and water, and more slowly anaerobically to β-hydroxybutyrate and acetoacetate. The evidence that poly-β-hydroxybutyrate is a reserve carbon and energy source is discussed.
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