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首页> 外文期刊>Separation and Purification Technology >Purification and identification of a novel cutinase from Coprinopsis cinerea by adsorptive bubble separation
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Purification and identification of a novel cutinase from Coprinopsis cinerea by adsorptive bubble separation

机译：吸附性气泡分离法纯化和鉴定灰葡萄粉新酶

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摘要

A novel extracellular esterase was separated from culture supernatants of the basidiomycete Coprinopsis cinerea by means of foam fractionation. The parameters pH value, gas flow rate, addition of detergents, and the column design were varied to optimise the transport of the active enzyme into the foam phase. On the 70-mL scale, a recovery of activity of 79% with an enrichment factor of 10.5 was obtained at pH 7 and 20 mL air min~(-1) within 15 min. The enriched enzyme was characterized biochemically by semi-native SDS-PAGE and IEF electrophoresis with activity staining. Peptide sequencing was performed after tryptic digestion by electrospray tandem mass spectrometry. Homology searches revealed significant similarities to cutinases of various ascomycetes and to C. cinerea genome data recently annotated as cutinases.
机译：通过泡沫分级分离将新的胞外酯酶与灰葡萄孢菌的培养上清液分离。改变参数pH值，气体流速，添加清洁剂和色谱柱设计，以优化活性酶向泡沫相的传输。在70 mL规模下，在pH 7和20 mL air min〜（-1）的情况下，在15分钟内获得了79％的活性回收率和10.5的富集因子。通过半天然SDS-PAGE和带有活性染色的IEF电泳对富集的酶进行生化鉴定。在胰蛋白酶消化后，通过电喷雾串联质谱法进行肽测序。同源性搜索显示与各种子囊囊角质酶和最近被注释为角质酶的灰质曲霉基因组数据具有显着相似性。

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