A novel esterase from a soil metagenomic library displaying a broad substrate range.
A novel esterase from a soil metagenomic library displaying a broad substrate range.
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来自土壤宏基因组文库的新型酯酶，具有广泛的底物范围
DOI：
10.1186/s13568-021-01198-5
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发表时间：
2021-03-05
期刊：
影响因子：
3.7
通讯作者：
Yin S
中科院分区：
文献类型：
--
作者：
Yao J;Gui L;Yin S
从土壤宏基因组文库中分离到一个新的酯酶基因。该基因编码一个520个氨基酸的蛋白质，其中含有一个21个氨基酸的信号肽。一级结构分析表明，该蛋白含有一个保守的活性位点基序（SxSxG）和一个结构基序（CS-D-HC）。在大肠杆菌BL 21（DE 3）中进行了酯酶基因的克隆和表达。纯化的酯酶的SDS-PAGE分析表明，它是在一个高度可溶性的形式表达，其分子量估计为55 kDa。该酯酶对短酰基对硝基苯酯，特别是对乙酸对硝基苯酯具有较高的活性，表明该酶是一种典型的羧酸酯酶，而不是脂肪酶。该酶以乙酸对硝基苯酯为底物，在pH7.0和30 °C下具有最佳活性，在pH4.5 ~ 10.0范围内稳定，温度不高于50 °C。此外，该酶对底物的专一性与已知的酯酶不同，它不仅能水解对硝基苯酯，而且能水解所有的羟基苯甲酸酯和羟基肉桂酸酯。由于它是一种对广泛的酚酯具有活性的酶，同时具有阿魏酸酯酶、绿原酸酯酶和单宁酶活性，因此它可以作为生物技术应用的有价值的候选物。
A novel esterase gene was isolated from a soil metagenomic library. The gene encoded a protein of 520 amino acids which contained a 21 aa signal peptide. Primary structure analysis of the protein sequence revealed that it contained a conserved active site motif (SxSxG) and a structural motif (CS-D-HC). Then the esterase gene was cloned and expressed inEscherichia coli BL21(DE3). SDS-PAGE analysis of the purified esterase showed that it was expressed in a highly soluble form and its molecular mass was estimated to be 55 kDa. Characterization of the esterase revealed that it exhibited high activity towardp-nitrophenyl esters with short acyl chains and especiallyp-nitrophenyl acetate, suggesting that it was a typical carboxylesterase rather than a lipase. Withp-nitrophenyl acetate as substrate, the enzyme showed its optimal activity at pH 7.0 and 30 °C, and it was stable at a broad pH range from 4.5 to 10.0 and temperature not higher than 50 °C. Furthermore, the enzyme showed different substrate specificity from known esterase, it was not only hydrolyzing againstp-nitrophenyl esters, but also hydrolyzing all hydroxybenzoic esters and hydroxycinnamic ester assayed. As it was an enzyme active on a broad range of phenolic esters, simultaneously possessing feruloyl esterase, chlorogenate esterase and tannase activities, it could serve as a valuable candidate for applications in biotechnology.
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影响因子：
4.1
作者：
Crepin, VF;Faulds, CB;Connerton, IF
通讯作者：
Connerton, IF
影响因子：
3
作者：
Ramirez, L.;Arrizon, J.;Mateos-Diaz, J. C.
通讯作者：
Mateos-Diaz, J. C.
影响因子：
4.4
作者：
Esteban-Torres, Maria;Maria Landete, Jose;Munoz, Rosario
通讯作者：
Munoz, Rosario
影响因子：
4.4
作者：
Esteban-Torres, Maria;Reveron, Ines;Munoz, Rosario
通讯作者：
Munoz, Rosario
影响因子：
5.8
作者：
Larkin, M. A.;Blackshields, G.;Higgins, D. G.
通讯作者：
Higgins, D. G.