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中国精品科技期刊2020
张杰,侯珺淇,代振玉,等. 一株产羧酸酯酶菌的鉴定及酯酶特征的研究[J]. 食品工业科技,2021,42(19):126−134. doi: 10.13386/j.issn1002-0306.2020120275.
引用本文: 张杰,侯珺淇,代振玉,等. 一株产羧酸酯酶菌的鉴定及酯酶特征的研究[J]. 食品工业科技,2021,42(19):126−134. doi: 10.13386/j.issn1002-0306.2020120275.
ZHANG Jie, HOU Junqi, DAI Zhenyu, et al. Identification of a Strain with Carboxylesterase and Its Enzymatic Character[J]. Science and Technology of Food Industry, 2021, 42(19): 126−134. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2020120275.
Citation: ZHANG Jie, HOU Junqi, DAI Zhenyu, et al. Identification of a Strain with Carboxylesterase and Its Enzymatic Character[J]. Science and Technology of Food Industry, 2021, 42(19): 126−134. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2020120275.

一株产羧酸酯酶菌的鉴定及酯酶特征的研究

Identification of a Strain with Carboxylesterase and Its Enzymatic Character

  • 摘要: 目的:鉴定中国白酒发酵过程中的微生物种类,研究关键酶的特征与作用机制有利于提高白酒的优质品率。方法:本研究用形态学和生理学、16S rRNA、gyr B基因和antiSMASH分析的方法对酒曲中的一株微生物进行了验证;对该菌株的特性进行了研究,采用分子建模的方法获得了羧酸酯酶的3D模型,用分子对接的方法探讨了该菌株的羧酸酯酶的机理。结果:该菌株为产羧酸酯酶的革兰氏阴性菌Pb1(MW580690);该菌株呈现典型的S型生长曲线,产物曲线为S型,羧酸酯酶活化最优pH范围为5.0~9.0。分子对接结果显示Phe21A为该酶具有催化活性的主要氨基酸,水解三丁酸甘油酯为丁酸和甘油。分子对接结果显示三丁酸甘油酯经过构象变化被转移到催化中心后,进一步被加工;酶的亲水性和疏水性的相互作用表面有利于配体向下转移,进而从疏水性通道释放产物到的酶表面。结论:产羧酸酯酶的菌株为贝莱斯芽胞杆菌,并为羧酸酯酶水解三丁酸甘油酯类物质的底物识别、转移和催化机理提供了新的见解。

     

    Abstract: Objective: Microbial species level identification in Chinese liquor fermentation, key enzymatic character and enzymatic mechanism study benefits to improve quality of Chinese liquor. Methods: Based on the morphological and physiological characteristics, 16S rRNA, gyrB gene and antiSMASH analysis to identify a micro-organism. 3D model by 3D structure modeling was gained and the mechanism regulating by molecular docking was insighted. Results: The strain was identified as gram-negative Bacillus velezensis with carboxylesterase gene. The growth curve was typical S-shaped growth curve. Product curve was typical S-shaped. The range of carboxylesterase-activity was from pH5.0 to pH9.0. Molecular docking indicated that Phe21A was responsible for the primarily hydrolysis reaction in catalytic site where glyceryl tributyrate was hydrolyzed into glycerol and butyric acid. Glyceryl tributyrate experiencing conformational changes were transported into catalytic site and then hydrolysed; Interactions of hydrophilic and hydrophobic interface were beneficial to the substrate transferred downward. The hydrolyzed substrates went to the enzyme surface from hydrophobic channel. Conclusion: The stain is identificated as gram-negative Bacillus velezensis with carboxylesterase gene and this study provides a new view of substrate recognition, transfer and catalysis mechanism when carboxylesterase hydrolyzes substances, such as glyceryl tributyrate.

     

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