kre1Δ 酿酒酵母孢子固定肌酐酶体系的构建及其酶学分析

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  • 1(江南大学 生物工程学院 糖化学与生物技术教育部重点实验室,江苏 无锡,214122)
博士研究生

网络出版日期: 2018-03-14

基金资助

国家自然科学基金(21778023,21576118)

Construction and analysis of creatininase encapsulated in kre1Δ Saccharomyces cerevisiae yeast spores

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  • PAN Hua-ping1,Nakanishi Hideki1,KONG Jun1,GAO Xiao-dong1*

Online published: 2018-03-14

摘要

酵母孢子固定化酶是一种利用酵母孢子对酶进行包埋固定的新型酶固定化技术。研究表明利用有孢子壁缺陷的osw2Δ酿酒酵母孢子作为酶固定化载体,不仅能大幅提高酶活,而且具有较强的抗逆性及较好的可重复利用性。为了研究新发现的kre1Δ孢子是否具有类似的性能,将肌酐酶转入酵母孢子,并对其酶学性能进行分析。实验结果表明kre1Δ孢子固定的肌酐酶活性与osw2Δ孢子相近,都显著高于野生型孢子。与游离酶相比,kre1Δ孢子固定的肌酐酶与osw2Δ一样,都有较好的可重复利用性和较强的抗逆性。同时,kre1Δ孢子与osw2Δ孢子肌酐酶活的最适温度均为60oC,最适pH值均为6.0,且kre1Δ孢子与osw2Δ孢子一样都表现出较好的耐高温性及较广范围的pH耐受性。因此,kre1Δ孢子也是综合性能较好的酶固定化载体。

本文引用格式

潘华平, 中西秀树, 孔军, 等 . kre1Δ 酿酒酵母孢子固定肌酐酶体系的构建及其酶学分析[J]. 食品与发酵工业, 2018 , 44(2) : 1 . DOI: 10.13995/j.cnki.11-1802/ts.015780

Abstract

Encapsulated enzymes by using yeast spores is a new enzyme-immobilized technology. It was reported that the activity of encapsulated enzymes was largely improved bySaccharomyces cerevisiaespores ofosw2Δwhich has mild spore wall defective and theosw2Δspore-encapsulate enzymes also exhibit resistance to environmental stresses and have a good reusability. To investigate whetherkre1Δspores have similar ability, the creatininase gene was transformed into yeast cells and the properties of encapsulated enzymes were examined. The results suggested the activity of the creatininase encapsulated inkre1Δspores was similar toosw2Δspores but was significantly higher than that of wild type spores. Compared with free enzymes, likeosw2Δ,encapsulated creatininase inkre1Δspores exhibits resistance to environmental stresses and has a good reusability. The spore-encapsulated creatininase inkre1Δandosw2Δshows highest activity at 60oC, pH 6.0. Moreover, thekre1Δspore-encapsulated enzymes are resistant to high temperature and exhibit a wide range of pH tolerance as same as that ofosw2Δspores. Therefore,kre1Δspores can be used as a superior carrier of enzyme capsules.
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