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-  2018 

约氏乳酸杆菌中一种新的阿魏酸酯酶基因的克隆表达与性质研究

DOI: 10.3969/j.issn.1673-1689.2018.11.002

Keywords: 阿魏酸酯酶 克隆表达 酶学性质 约氏乳酸杆菌
ferulic acid esterase
,cloning and expression,enzymatic property, Lactobacillus johnsonii

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Abstract:

阿魏酸是一种重要的抗氧化剂,是近几年研究的热点。阿魏酸酯酶(FAEs)能够水解酯键来释放游离的阿魏酸。本文作者根据约氏乳酸杆菌(Lactobacillus johnsonii)的全基因组序列和阿魏酸酯酶保守序列设计了一个新的阿魏酸酯酶的编码基因Lj0900。通过PCR扩增,酶切连接,成功构建重组载体,并在大肠杆菌(BL21)中成功表达。对表达的FAEs用镍柱进行纯化,对获得纯酶进行了初步的酶学性质研究,用阿魏酸甲酯为底物的情况下,发现该酶的最适温度为30 ℃,最适pH 6。
Ferulic acid is an important antioxidant,is a hot topic research in recent years. Ferulic acid esterase(FAEs) can hydrolyze the ester bond to release free ferulic acid. In this paper,according to the whole genome sequence of Lactobacillus johnsonii and the conserved sequence of ferulic acid esterase,the author designed a new encoding gene Lj0900 of ferulic acid esterase. By PCR amplification,enzyme connection,constructing the restructuring of the carrier,and successfully expressed in E. coli BL21. To study the preliminary properties of pure enzyme,crude FAEs was purified by nickel column. With the methyl ferulate as the substrate,discovered that the optimum temperature for the enzyme is 30 ℃,the optimum pH is 6

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