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Purification and preliminary characterization of the extracellular lipase of Bacillus subtilis 168, an extremely basic pH-tolerant enzyme.

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Lesuisse, E. [UCL] Colson, Charles [UCL] Schanck, K. [UCL]

The extracellular lipase of Bacillus subtilis 168 was purified from the growth medium of an overproducing strain by ammonium sulfate precipitation followed by phenyl-Sepharose and hydroxyapatite column chromatography. The purified lipase had a strong tendency to aggregate. It exhibited a molecular mass of 19,000 Da by SDS-PAGE and a pI of 9.9 by chromatofocusing. The enzyme showed maximum stability at pH 12 and maximum activity at pH 10. The lipase was active toward p-nitrophenyl esters and triacylglycerides with a marked preference for esters with C8 acyl groups. Using trioleyl glycerol as substrate, the enzyme preferentially cleaved the 1(3)-position ester bond. No interfacial activation effect was observed with triacetyl glycerol as substrate.

Document type Article de périodique (Journal article) – Journal Article, Research Support, Non-U.S. Gov't
Publication date 1993
Journal information "European journal of biochemistry / FEBS" - Vol. 216, no. 1, p. 155-60 (1993)
Peer reviewed yes
issn 0014-2956
Publication status Publié
Affiliation UCL - MD/MIGE - Département de microbiologie, d'immunologie et de génétique
MESH Subject Ammonium Sulfate ; Bacillus subtilis - enzymology ; Calcium - pharmacology ; Chromatography, Thin Layer ; Durapatite ; Electrophoresis, Polyacrylamide Gel ; Enzyme Activation ; Hydrogen-Ion Concentration ; Hydroxyapatites ; Lipase - chemistry - isolation & purification - metabolism ; Molecular Weight ; Phenylmethylsulfonyl Fluoride - pharmacology ; Substrate Specificity ; Temperature ; Triglycerides - metabolism
Links
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Bibliographic reference Lesuisse, E. ; Colson, Charles ; Schanck, K.. Purification and preliminary characterization of the extracellular lipase of Bacillus subtilis 168, an extremely basic pH-tolerant enzyme.. In: European journal of biochemistry / FEBS, Vol. 216, no. 1, p. 155-60 (1993)
Permanent URL http://hdl.handle.net/2078.1/12096