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从藤黄微球菌中一步法纯化不同的脂肪酶。

Single-step purification of different lipases from Staphylococcus warneri.

机构信息

Departamento de Biocatalisis, Instituto de Catalisis (CSIC), Madrid, Spain.

出版信息

J Chromatogr A. 2010 Jan 22;1217(4):473-8. doi: 10.1016/j.chroma.2009.11.055. Epub 2009 Nov 20.

Abstract

Three different lipases from the extract crude of Staphylococcus warneri have been purified by specific lipase-lipase interactions using different lipases (TLL, RML, PFL, BTL2) covalently attached to a solid support as adsorption matrix. BTL2 immobilized on glyoxyl-DTT adsorbed selectivity only a 30 kDa lipase from the crude, which was desorbed by adding 0.1% triton X-100. Using glyoxyl-PFL as matrix, two new lipases (28 and 40 kDa) were adsorbed, and completely pure 40 kDa lipase was obtained after desorption using 0.01% triton, whereas 28 kDa lipase was desorbed after the incubation of the lipase matrix with 3% detergent. When using other matrixes as glyoxyl-TLL or glyoxyl-RML, different lipases were adsorbed. This methodology could be a very efficient and useful method to purify several lipases from crude extracts from different sources.

摘要

已使用不同的固定化脂肪酶(TLL、RML、PFL、BTL2)作为吸附基质,通过特定的脂肪酶-脂肪酶相互作用,从金黄色葡萄球菌粗提物中纯化出三种不同的脂肪酶。BTL2 固定在乙二醛-DTT 上,仅选择性吸附粗提物中的一种 30 kDa 脂肪酶,该酶可通过添加 0.1% Triton X-100 洗脱。使用乙二醛-PFL 作为基质,吸附了两种新的脂肪酶(28 和 40 kDa),经 0.01% Triton 洗脱后可获得纯 40 kDa 脂肪酶,而 28 kDa 脂肪酶在与 3%洗涤剂孵育后被洗脱。当使用其他基质(如乙二醛-TLL 或乙二醛-RML)时,吸附了不同的脂肪酶。该方法可能是一种非常有效和有用的方法,可从不同来源的粗提物中纯化多种脂肪酶。

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