The Isolation and Estimation of the Poly- -hydroxy-butyrate Inclusions of Bacillus Species

SUMMARY: 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.