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A number of detergents will damage the lipoproteins of the microbial cell membrane and lead to release of intracellular components. The compounds which can

Fig. 10.21. Details of homogonizer valve assembly (Brookman, 1974). (A) 0-50,000 psi pressure transducer; (B) pressure-control handwheel; (C) linear variable displacement transformer; (->) direction of flow.

be used for this purpose include quaternary ammonium compounds, sodium lauryl sulphate, sodium dodecyl sulphate (SDS) and Triton X-100. Unfortunately, the detergents may cause some protein denaturation and may need to be removed before further purification stages can be undertaken. Pullulanase is an enzyme which is bound to the outer membrane of Klebsiella pneumoniae. The cells were suspended in pH 7.8 buffer and 1% sodium cholate was added. The mixture was stirred for 1 hour to solubilize most of the enzyme (Kroner et al., 1978). The use of Triton X-100 in combination with guanidine-HCl is widely and effectively used for the release of cellular protein (Naglak and Wang, 1992; Hettwer and Wang, 1989), Hettwer and Wang obtaining greater than 75% protein release in less than one hour from Escherichia coli under fermentation conditions.

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