Identification of a Strain with Carboxylesterase and Its Enzymatic Character

ZHANG Jie; HOU Junqi; DAI Zhenyu; ZHAO Xin; HOU Xiaoge

doi:10.13386/j.issn1002-0306.2020120275

Volume 42 Issue 19

Sep. 2021

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Science and Technology of Food Industry > 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.

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Identification of a Strain with Carboxylesterase and Its Enzymatic Character

College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou 466001, China

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Received Date: January 03, 2021
Available Online: August 10, 2021

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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.
- 16S rRNA,
- carboxylesterase,
- molecular docking,
- Bacillus velezensis

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References

[1]	赖登燡, 范威. 中国白酒与威士忌酒的典型性对比研究[J]. 酿酒科技,2007(8):155−157. [Lai Dengyi, Fan Wei. Study on the typicality of chinese liquor and whiskey[J]. Liquor-making Science & Technology,2007(8):155−157. doi: 10.3969/j.issn.1001-9286.2007.08.047
[2]	Jin G, Zhu Y, Xu Y. Mystery behind Chinese liquor fermentation[J]. Trends Food Sci Technol,2017,63:18−28. doi: 10.1016/j.jpgs.2017.02.016
[3]	黄丹, 张强, 严芳, 等. 浓香型大曲中酯化酶细菌的分离鉴定及产酶条件研究[J]. 中国酿造,2009(8):57−59. [Huang Dan, Zhang Qiang, Yan Fang, et al. Isolation and identification of esterifying enzyme-producing bacteria from Luzhou-flavor Daqu and related enzyme production conditions[J]. China Brewing,2009(8):57−59. doi: 10.3969/j.issn.0254-5071.2009.08.016
[4]	李绍亮, 李亚凤, 李学思. 酿酒大曲中酯化红曲的分离及应用[J]. 酿酒,2014,41(2):32−39. [Li Shaoliang, Li Yafeng, LI Xuesi. Isolation and application of esterifying Monascus in the liquor making Koji[J]. Liquor Making,2014,41(2):32−39. doi: 10.3969/j.issn.1002-8110.2014.02.010
[5]	刘建文, 吴生文, 朱力红, 等. 特香型大曲中纤维素酶、半纤维素酶、蛋白酶、酯化酶活力变化规律的研究[J]. 酿酒,2013,40(5):40−42. [Liu Jianwen, Wu Shengwen, Zhu Lihong, et al. Investigate on the cellulase and hemicellulose and protease and esterifying enzyme activity of the site-flavor Daqu[J]. Liquor Making,2013,40(5):40−42. doi: 10.3969/j.issn.1002-8110.2013.05.012
[6]	胡晓龙, 赵东, 葛隐, 等. 五粮液大曲中产酯化酶红曲霉的分离及酯化酶的纯化[J]. 酿酒,2011,38(3):35−8. [Hu Xiaolong, Zhao Dong, Ge Yin, et al. The isolation of esterified Monascus from Wuliangye Daqu and purification of esterifying enzyme[J]. Liquor Making,2011,38(3):35−8. doi: 10.3969/j.issn.1002-8110.2011.03.012
[7]	刘阳, 薛正莲, 黄祖耀, 等. 中高温大曲中产酯酶细菌的筛选[J]. 中国酿造,2012,31(8):99−102. [Liu Yang, Xue Zhenglian, Huang Zuyao, et al. Screening of esterase-producing bacteria from medium and high temperature Daqu[J]. China Brewing,2012,31(8):99−102. doi: 10.3969/j.issn.0254-5071.2012.08.027
[8]	滕巍, 李国莹, 刘小波, 等. 大曲中产酯化酶菌株的分离鉴定及固体发酵工艺优化[J]. 食品与生物技术学报,2016,35(9):971−977. [Teng Wei, Li Guoying, Liu Xiaobo, et al. Isolation and identification of ethyl caproate esterifying enzyme in Daqu and its fermentative technology optimization[J]. Journal of Food Science and Biotechnology,2016,35(9):971−977. doi: 10.3969/j.issn.1673-1689.2016.09.011
[9]	侯小歌, 张杰, 孙忠科, 等. 大曲产酯化酶芽孢菌的分离鉴定及其发酵条件优化研究[J]. 轻工学报,2018,33(4):13−20. [Hou Xiaoge, Zhang Jie, Sun Zhongke, et al. Isolation, identification and fermentation conditions optimization of an esterase-producing Bacillus strain from Daqu[J]. Journal of Light Industry,2018,33(4):13−20. doi: 10.3969/j.issn.2096-1553.2018.04.002
[10]	唐取来, 李晶晶, 郭学武, 等. 清香型大曲酯化酶活力的研究[J]. 中国酿造,2017,36(1):35−38. [Tang Qulai, Li Jingjing, Guo Xuewu, et al. Esterifying enzymes activity of light-flavor Daqu[J]. China Brewing,2017,36(1):35−38.
[11]	黄建华, 张煜行, 张希君, 等. 金属离子对老白干酒曲酯化酶的影响[J]. 中国酿造,2007,169(4):24−26. [Huang Jianhua, Zhang Yuhang, Zhang Xijun, et al. Preliminary research on the impact of metal ions on esterification enzyme in laobaigan liquor qu[J]. China Brewing,2007,169(4):24−26. doi: 10.3969/j.issn.0254-5071.2007.04.007
[12]	Miller G L, Blum R, Glennon W E, et al. Measurement of carboxymethylcellulase activity[J]. AnBio,1960,1(2):127−132.
[13]	Tamura K, Dudley J, Nei M, et al. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Mol Biol Evol,2007,24(8):1596−1599. doi: 10.1093/molbev/msm092
[14]	Bailey T L, Boden M, Buske F A, et al. MEME Suite: tools for motif discovery and searching[J]. Nucleic Acids Research,2009,37:202−208. doi: 10.1093/nar/gkp335
[15]	Cho M S, Jin Y J, Kang B K, et al. Understanding the ontogeny and succession of Bacillus velezensis and B. subtilis subsp. subtilis by focusing on kimchi fermentation[J]. Sci Rep,2018,8(1):7045−7048. doi: 10.1038/s41598-018-25514-5
[16]	Laskowski R A, Swindells M B. LigPlot+: multiple ligand-protein interaction diagrams for drug discovery[J]. J Chem Inf Model,2011,51(10):2778−2786. doi: 10.1021/ci200227u
[17]	Chen Meichun, Wang Jiangping, Liu Bo, et al. Biocontrol of tomato bacterial wilt by the new strain Bacillus velezensis FJAT-46737 and its lipopeptides[J]. BMC microbiology,2020,20(1):160−172. doi: 10.1186/s12866-020-01851-2
[18]	Chen C, Chen H, Zhang Y, et al. TBtools: An Integrative toolkit developed for interactive analyses of big biological data[J]. Molecular Plant,2020,13(8):1194−1202. doi: 10.1016/j.molp.2020.06.009
[19]	Bienert S, Waterhouse A, De Beer Tjaart A P, et al. The SWISS-MODEL Repository-new features and functionality[J]. Nucleic Acids Research,2017,45:313−319. doi: 10.1093/nar/gkw1132
[20]	Waterhouse A, Bertoni M, Bienert S, et al. SWISS-MODEL: homology modelling of protein structures and complexes[J]. Nucleic Acids Research,2018,46:296−303.
[21]	Schwede T, Haas J, Gumienny R, et al. QMEANDisCo-distance constraints applied on model quality estimation[J]. Bioinformatics,2020,36(6):1765−1771. doi: 10.1093/bioinformatics/btz828
[22]	Chen Z, Zhang X, Peng C, et al. D3Pockets: A method and web server for systematic analysis of protein pocket dynamics[J]. J Chem Inf Model,2019,59(8):3353−3358. doi: 10.1021/acs.jcim.9b00332
[23]	Pettersen E F, Goddard T D, Huang C C, et al. UCSF Chimera--a visualization system for exploratory research and analysis[J]. J Comput Chem,2004,25(13):1605−1612. doi: 10.1002/jcc.20084
[24]	张浩, 胡智慧, 刑爽, 等. 酸浓度和pH值对浓香型大曲酯化酶催化活力的影响[J]. 中国酿造,2017,36(5):41−45. [Zhang Hao, Hu Zhihui, Xing Shuang, et al. Effect of acid concentration and pH on esterifying enzyme catalytic activity of Luzhou-flavor Daqu[J]. China Brewing,2017,36(5):41−45. doi: 10.11882/j.issn.0254-5071.2017.05.009
[25]	Fan B, Blom J, Klenk H, et al. Bacillus amyloliquefaciens, Bacillusvelezensis, and form an "Operational Group " within the species complex[J]. Frontiers in microbiology,2017,8(22):1−15.
[26]	Madhaiyan M, Poonguzhali S, Kwon S, et al. Bacillus methylotrophicus sp. nov., a methanol-utilizing, plant-growth-promoting bacterium isolated from rice rhizosphere soil[J]. Int J Syst Evol Microbiol,2010,60:2490−2495. doi: 10.1099/ijs.0.015487-0
[27]	Robert X, Gouet P. Deciphering key features in protein structures with the new ENDscript server[J]. Nucleic acids research,2014,42:320−3244.
[28]	沈怡方. 白酒中四大乙酯在酿造发酵中形成的探讨[J]. 酿酒科技,2003(5):28−31. [Shen Yifang. Study on the formation mechanism of four main kinds of ethyl esters in the fermentation of liquors[J]. Liquor-making Science & Technology,2003(5):28−31. doi: 10.3969/j.issn.1001-9286.2003.05.006
[29]	Arnold K, Bordoli L, Kopp J, et al. The SWISS-MODEL workspace: A web-based environment for protein structure homology modeling[J]. Bioinformatics,2006,22:195−201. doi: 10.1093/bioinformatics/bti770
[30]	Zhu J, Chen C, Zhang B, et al. The inhibitory effects of flavonoids on α-amylase and α-glucosidase[J]. Crit Rev Food Sci Nutr,2019,60(4):695−708.
[31]	Silvestrini L, Cianci M. Principles of lipid-enzyme interactions in the limbus region of the catalytic site of Candida antarctica lipase B[J]. Int J Biol Macromol,2020,158:358−363. doi: 10.1016/j.ijbiomac.2020.04.061