传统食品发酵环境宏基因组中酯酶基因的克
发布时间：2019-06-06 05:53
【摘要】：微生物酯酶是一种广泛应用于食品、医药和精细化工等领域的工业化酶。为了丰富酯酶资源,本文通过提取我国传统发酵食品环境样品总基因组DNA,构建宏基因组文库,从中筛选获得一个新的酯酶基因(est_115),该基因全长948 bp,编码316个氨基酸,其蛋白序列同源性分析,表明与来自Pseudomonas lutea的羧酸酯水解酶同源性最高为38%,显示酯酶Est_115属于新的酯酶类。然后构建了est_115基因重组表达载体,在大肠杆菌BL21(DE3)中获得表达,经纯化得到重组酯酶Est_115。酶学性质的分析表明,该重组酯酶Est_115对酰基碳链较短的对硝基苯酚酯具有较高的催化活性,最适作用温度为35℃,最适p H值为7.0,在10%~18%Na Cl条件下,酶蛋白可保持较高的催化活性,具有良好的耐盐性,提示该酶可以应用在高渗透压食品加工中,是一个新型的极具潜力的酯酶。
[Abstract]:Microbial esterase is an industrial enzyme widely used in food, medicine and fine chemical industry. In order to enrich esterase resources, a new esterase gene (est_115) was obtained by extracting total genomic DNA, from traditional fermented food environmental samples in China to construct macrogenomic library. The full length of the gene was 948 bp,. It encodes 316 amino acids and its protein sequence homology analysis shows that the homology with Carboxyester hydrolase from Pseudomonas lutea is up to 38%, indicating that esterase Est_115 belongs to a new esterase. Then the recombinant expression vector of est_115 gene was constructed and expressed in E. coli BL21 (DE3). The recombinant esterase Est_115. was purified. The analysis of enzyme properties showed that the recombinant esterase Est_115 had high catalytic activity for p-nitrophenol ester with short acyl carbon chain, the optimum reaction temperature was 35 鈩，
本文编号：2494115
[Abstract]:Microbial esterase is an industrial enzyme widely used in food, medicine and fine chemical industry. In order to enrich esterase resources, a new esterase gene (est_115) was obtained by extracting total genomic DNA, from traditional fermented food environmental samples in China to construct macrogenomic library. The full length of the gene was 948 bp,. It encodes 316 amino acids and its protein sequence homology analysis shows that the homology with Carboxyester hydrolase from Pseudomonas lutea is up to 38%, indicating that esterase Est_115 belongs to a new esterase. Then the recombinant expression vector of est_115 gene was constructed and expressed in E. coli BL21 (DE3). The recombinant esterase Est_115. was purified. The analysis of enzyme properties showed that the recombinant esterase Est_115 had high catalytic activity for p-nitrophenol ester with short acyl carbon chain, the optimum reaction temperature was 35 鈩，
本文编号：2494115
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