A Protease-Insensitive Feruloyl Esterase from China Holstein Cow Rumen Metagenomic Library: Expression, Characterization, and Utilization in Ferulic Acid Release from Wheat Straw
A Protease-Insensitive Feruloyl Esterase from China Holstein Cow Rumen Metagenomic Library: Expression, Characterization, and Utilization in Ferulic Acid Release from Wheat Straw
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DOI：
10.1021/jf204556u
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发表时间：
2012-03-14
影响因子：
6.1
通讯作者：
Shen, Lin
中科院分区：
文献类型：
--
作者：
Cheng, Fansheng;Sheng, Jiping;Shen, Lin
建立中国荷斯坦奶牛瘤胃微生物元基因组文库，筛选新基因簇。鉴定出一个新的阿铁酰酯酶（FAE）基因，全长789 bp，其编码的蛋白与已知酯酶序列的同源性为56%。该基因在大肠杆菌BL21 （DE3）中功能表达，重组蛋白总分子量为32.4 kDa。纯化后的酶对羟基肉桂酸的四种甲酯具有广泛的特异性，在pH 8.0, 40℃的最佳条件下对阿魏酸甲酯具有较高的活性（259.5 U/mg）。还观察到高的热稳定性和pH稳定性。此外，该酶对蛋白酶表现出广泛的抗性。FAE-SH1能通过纤维素酶、β -1,4-内木聚糖酶、β -1,3-葡聚糖酶和果胶酶促进麦草阿魏酸的释放。这些特征表明FAE-SH1是促进生物质降解和改善食品和饲料健康效应的良好候选者。
A metagenomic library of China Holstein cow rumen microbes was constructed and screened for novel gene cluster. A novel feruloyl esterase (FAE) gene was identified with a length of 789 bp and encoded a protein displaying 56% identity to known esterase sequences. The gene was functionally expressed in Escherichia coli BL21 (DE3), and the total molecular weight of the recombined protein was 32.4 kDa. The purified enzyme showed a broad specificity against the four methyl esters of hydroxycinnamic acids and high activity (259.5 U/mg) to methyl ferulate at optimum conditions (pH 8.0, 40 degrees C). High thermal and pH stability were also observed. Moreover, the enzyme showed broad resistance to proteases. FAE-SH1 can enhance the release of ferulic acid from wheat straw with cellulase, beta-1,4-endoxylanase, beta-1,3-glucanase, and pectase. These features suggest FAE-SH1 as a good candidate to enhance biomass degradation and improve the health effects of food and forage.