QUANTITATIVE SPECTROPHOTOMETRIC ASSAY FOR STAPHYLOCOCCAL LIPASE

被引:55
作者
SMELTZER, MS [1 ]
HART, ME [1 ]
IANDOLO, JJ [1 ]
机构
[1] KANSAS STATE UNIV AGR & APPL SCI,COLL VET MED,DEPT PATHOL & MICROBIOL,MANHATTAN,KS 66506
关键词
D O I
10.1128/AEM.58.9.2815-2819.1992
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We report the development of a specific spectrophotometric assay for the quantitative determination of lipase activity in Staphylococcus aureus. The assay is based on the rate of clearance of a tributyrin emulsion, and it can detect as little as 1.0-mu-g of purified Pseudomonas lipase per ml. By comparison with the reaction rates obtained with Pseudomonas lipase, we calculated that S. aureus PS54C and S6C produce approximately 15 and 60-mu-g of extracellular lipase per ml, respectively. Neither PS54, which is lysogenized with the converting bacteriophage L54a and is consequently lipase negative (Lip-), nor KSI905, a Lip- transpositional mutant of strain S6C, was positive in our spectrophotometric assay. The specificity of the spectrophotometric tributyrin assay was confirmed with a triolein plate assay; supernatants from S6C and PS54C hydrolyzed triolein, while supernatants from PS54 and KSI905 did not. In contrast to the results of the spectrophotometric tributyrin assay, all enzyme preparations tested (including commercially purified esterase) were positive when examined by a tributyrin plate assay. The lack of specificity in the tributyrin plate assay emphasizes the need to interpret the results of tributyrin lipolysis kinetically for assessing lipase activity in S. aureus.
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页码:2815 / 2819
页数:5
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