Identification and characterization of a novel β-glucosidase via metagenomic analysis of Bursaphelenchus xylophilus and its microbial flora.
Identification and characterization of a novel β-glucosidase via metagenomic analysis of Bursaphelenchus xylophilus and its microbial flora.
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通过松材线虫及其微生物菌群的宏基因组分析鉴定和表征新型β-葡萄糖苷酶
DOI：
10.1038/s41598-017-14073-w
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发表时间：
2017-11-01
影响因子：
4.6
通讯作者：
Niu Q
中科院分区：
文献类型：
--
作者：
Zhang L;Fu Q;Li W;Wang B;Yin X;Liu S;Xu Z;Niu Q
β-葡糖苷酶催化纤维素水解的最后一步，并且在纤维素降解中是必需的。通过功能筛选，从松材线虫宏基因组文库中克隆到一个β-葡萄糖苷酶基因cen 502。分析表明，cen 502编码一个465个氨基酸的多肽，含有属于糖苷水解酶家族1（GH 1）的催化结构域。Cen 502异源表达，纯化和生化特征。重组Cen 502在pH 8.0和38 °C下显示最佳酶活性。该酶对对硝基苯基β D吡喃葡萄糖苷（pNPG; 180.3U/mg）的比活力最高，Km和Vmax分别为2.334 mol/ml和9.017 μmol/min/mg。Fe ~（2+）和Mn ~（2+）使Cen 502 β-葡萄糖苷酶活性分别提高60%和50%; Pb ~（2+）和K ~+使Cen 502 β-葡萄糖苷酶活性分别降低10%和25%。Cen 502表现出对多种底物的活性，包括纤维二糖、乳糖、水杨苷、地衣多糖、海带多糖和槐糖。然而，Cen 502显示出对β-1，4糖苷键而不是β-1，3、β-1，6或β-1，2键的水解的偏好。结果表明Cen 502是一种新的β-葡萄糖苷酶，来源于与B相关的细菌。木霉菌的纤维素酶，并可能代表一个有前途的目标，以提高效率的纤维素生物降解在工业应用中。
β-glucosidases catalyze the final step of cellulose hydrolysis and are essential in cellulose degradation. A β-glucosidase gene, cen502, was identified and isolated from a metagenomic library from Bursaphelenchus xylophilus via functional screening. Analyses indicated that cen502 encodes a 465 amino acid polypeptide that contains a catalytic domain belonging to the glycoside hydrolase family 1 (GH1). Cen502 was heterologously expressed, purified, and biochemically characterized. Recombinant Cen502 displayed optimum enzymatic activity at pH 8.0 and 38 °C. The enzyme had highest specific activity to p-nitrophenyl-β-D-glucopyranoside (pNPG; 180.3 U/mg) and had K m and V max values of 2.334 mol/ml and 9.017 μmol/min/mg, respectively. The addition of Fe2+ and Mn2+ significantly increased Cen502 β-glucosidase activity by 60% and 50%, respectively, while 10% and 25% loss of β-glucosidase activity was induced by addition of Pb2+ and K+, respectively. Cen502 exhibited activity against a broad array of substrates, including cellobiose, lactose, salicin, lichenan, laminarin, and sophorose. However, Cen502 displayed a preference for the hydrolysis of β-1,4 glycosidic bonds rather than β-1,3, β-1,6, or β-1,2 bonds. Our results indicate that Cen502 is a novel β-glucosidase derived from bacteria associated with B. xylophilus and may represent a promising target to enhance the efficiency of cellulose bio-degradation in industrial applications.
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影响因子：
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作者：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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