Sijia Liu , Yan Qin , Qingyan Wang , Jing Zhang , Jin Zhou , Baoxiang He , Xinquan Liang , Liang Xian , Junhua Wu
{"title":"来自芽孢杆菌 B58-2 的新型高特异性果胶酸裂解酶：基因克隆、异源表达和在苎麻脱胶中的应用","authors":"Sijia Liu , Yan Qin , Qingyan Wang , Jing Zhang , Jin Zhou , Baoxiang He , Xinquan Liang , Liang Xian , Junhua Wu","doi":"10.1016/j.enzmictec.2024.110395","DOIUrl":null,"url":null,"abstract":"<div><p><span><span>Pectinase plays a crucial role in </span>ramie<span> degumming. A gene encoding a putative pectate lyase from </span></span><span><em>Bacillus</em></span> sp. strain B58–2 was cloned and heterologously expressed in <em>Escherichia coli</em>. The amplified gene <em>BvelPL1</em><span> encoded a mature protein of 400 amino acids<span>. BvelPL1 shared the highest amino acid sequence identity (78.75%) with the enzymatically characterized pectate lyase Pel from </span></span><span><em>Bacillus subtilis</em></span><span><span> strain RCK (GenBank: AFH66771.1). The purified recombinant enzyme rBvelPL1-Ec exhibited a maximum specific activity of 2433.26 U/mg at pH 8.5 and 50 °C towards polygalacturonic acid. This specific activity was higher than that of most reported pectate lyases. Remarkably, the </span>enzymatic activity<span> of rBvelPL1-Ec increased by 23.28 times in the presence of 0.4 mM calcium ion<span>. The effect of calcium ion on promoting the enzymatic activity of rBvelPL1-Ec was greater than that for all reported pectate lyases. After degumming with rBvelPL1-Ec, a weight loss of 21.27 ± 1.17% of circled ramie fibers was obtained, and the surfaces of the ramie fibers became smoother. Moreover, a weight loss of 30.47 ± 0.46% was obtained through enzymatic treated and subsequent NaOH treated circled ramie fibers. The excellent performance in degumming suggests that rBvelPL1-Ec may serve as a promising biocatalyst in the textile industry.</span></span></span></p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"175 ","pages":"Article 110395"},"PeriodicalIF":3.4000,"publicationDate":"2024-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"A novel pectate lyase with high specific activity from Bacillus sp. B58-2: Gene cloning, heterologous expression and use in ramie degumming\",\"authors\":\"Sijia Liu , Yan Qin , Qingyan Wang , Jing Zhang , Jin Zhou , Baoxiang He , Xinquan Liang , Liang Xian , Junhua Wu\",\"doi\":\"10.1016/j.enzmictec.2024.110395\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><span><span>Pectinase plays a crucial role in </span>ramie<span> degumming. A gene encoding a putative pectate lyase from </span></span><span><em>Bacillus</em></span> sp. strain B58–2 was cloned and heterologously expressed in <em>Escherichia coli</em>. The amplified gene <em>BvelPL1</em><span> encoded a mature protein of 400 amino acids<span>. BvelPL1 shared the highest amino acid sequence identity (78.75%) with the enzymatically characterized pectate lyase Pel from </span></span><span><em>Bacillus subtilis</em></span><span><span> strain RCK (GenBank: AFH66771.1). The purified recombinant enzyme rBvelPL1-Ec exhibited a maximum specific activity of 2433.26 U/mg at pH 8.5 and 50 °C towards polygalacturonic acid. This specific activity was higher than that of most reported pectate lyases. Remarkably, the </span>enzymatic activity<span> of rBvelPL1-Ec increased by 23.28 times in the presence of 0.4 mM calcium ion<span>. The effect of calcium ion on promoting the enzymatic activity of rBvelPL1-Ec was greater than that for all reported pectate lyases. After degumming with rBvelPL1-Ec, a weight loss of 21.27 ± 1.17% of circled ramie fibers was obtained, and the surfaces of the ramie fibers became smoother. Moreover, a weight loss of 30.47 ± 0.46% was obtained through enzymatic treated and subsequent NaOH treated circled ramie fibers. The excellent performance in degumming suggests that rBvelPL1-Ec may serve as a promising biocatalyst in the textile industry.</span></span></span></p></div>\",\"PeriodicalId\":11770,\"journal\":{\"name\":\"Enzyme and Microbial Technology\",\"volume\":\"175 \",\"pages\":\"Article 110395\"},\"PeriodicalIF\":3.4000,\"publicationDate\":\"2024-01-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Enzyme and Microbial Technology\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0141022924000024\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Enzyme and Microbial Technology","FirstCategoryId":"5","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0141022924000024","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
A novel pectate lyase with high specific activity from Bacillus sp. B58-2: Gene cloning, heterologous expression and use in ramie degumming
Pectinase plays a crucial role in ramie degumming. A gene encoding a putative pectate lyase from Bacillus sp. strain B58–2 was cloned and heterologously expressed in Escherichia coli. The amplified gene BvelPL1 encoded a mature protein of 400 amino acids. BvelPL1 shared the highest amino acid sequence identity (78.75%) with the enzymatically characterized pectate lyase Pel from Bacillus subtilis strain RCK (GenBank: AFH66771.1). The purified recombinant enzyme rBvelPL1-Ec exhibited a maximum specific activity of 2433.26 U/mg at pH 8.5 and 50 °C towards polygalacturonic acid. This specific activity was higher than that of most reported pectate lyases. Remarkably, the enzymatic activity of rBvelPL1-Ec increased by 23.28 times in the presence of 0.4 mM calcium ion. The effect of calcium ion on promoting the enzymatic activity of rBvelPL1-Ec was greater than that for all reported pectate lyases. After degumming with rBvelPL1-Ec, a weight loss of 21.27 ± 1.17% of circled ramie fibers was obtained, and the surfaces of the ramie fibers became smoother. Moreover, a weight loss of 30.47 ± 0.46% was obtained through enzymatic treated and subsequent NaOH treated circled ramie fibers. The excellent performance in degumming suggests that rBvelPL1-Ec may serve as a promising biocatalyst in the textile industry.
期刊介绍：
Enzyme and Microbial Technology is an international, peer-reviewed journal publishing original research and reviews, of biotechnological significance and novelty, on basic and applied aspects of the science and technology of processes involving the use of enzymes, micro-organisms, animal cells and plant cells.
We especially encourage submissions on:
Biocatalysis and the use of Directed Evolution in Synthetic Biology and Biotechnology
Biotechnological Production of New Bioactive Molecules, Biomaterials, Biopharmaceuticals, and Biofuels
New Imaging Techniques and Biosensors, especially as applicable to Healthcare and Systems Biology
New Biotechnological Approaches in Genomics, Proteomics and Metabolomics
Metabolic Engineering, Biomolecular Engineering and Nanobiotechnology
Manuscripts which report isolation, purification, immobilization or utilization of organisms or enzymes which are already well-described in the literature are not suitable for publication in EMT, unless their primary purpose is to report significant new findings or approaches which are of broad biotechnological importance. Similarly, manuscripts which report optimization studies on well-established processes are inappropriate. EMT does not accept papers dealing with mathematical modeling unless they report significant, new experimental data.
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