Alginate as a macroaffinity ligand and an additive for enhanced activity and thermostability of lipases

被引:36
作者
Sharma, S [1 ]
Gupta, MN [1 ]
机构
[1] Indian Inst Technol, Dept Chem, New Delhi 110016, India
关键词
affinity precipitation; Chromobacterium lipase; porcine pancreatic lipase; protein purification; wheatgerm lipase;
D O I
10.1042/BA20000093
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Lipases are being employed increasingly for the synthesis of drug intermediates and pharmaceutically important molecules as well as for the resolution of racemic mixtures for obtaining physiologically active enantiomers. Alginate was used as a macroaffinity ligand to purify lipases from Chromobacterium viscosum, porcine pancreas and wheatgerm by employing the technique of affinity precipitation. In all the cases, the purified preparation showed a single band on SDS/PAGE. The process gave adequate yields of 87, 75 and 62% in the case of Chromobocterium, porcine and wheatgerm lipases respectively. Alginate was also found to activate the enzymes; this effect was most dramatic in the case of wheatgerm lipase, for which a 4-5-fold activity increase was observed. Furthermore, alginate was also found to protect the enzyme against thermoinactivation.
引用
收藏
页码:161 / 165
页数:5
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