A novel pectate lyase with high specific activity from Bacillus sp. B58-2: Gene cloning, heterologous expression and use in ramie degumming

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology Pub Date : 2024-01-16 DOI:10.1016/j.enzmictec.2024.110395

Sijia Liu , Yan Qin , Qingyan Wang , Jing Zhang , Jin Zhou , Baoxiang He , Xinquan Liang , Liang Xian , Junhua Wu

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引用次数: 0

Abstract

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.

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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

果胶酶在苎麻脱胶过程中起着至关重要的作用。克隆并在大肠杆菌中异源表达了来自芽孢杆菌 B58-2 菌株的果胶酶基因。扩增的基因 BvelPL1 可编码 400 个氨基酸的成熟蛋白质。BvelPL1 与经酶切鉴定的枯草芽孢杆菌 RCK 菌株的果胶酸裂解酶 Pel（GenBank：AFH66771.1）氨基酸序列相同度最高（78.75%）。纯化的重组酶 rBvelPL1-Ec 在 pH 值为 8.5、温度为 50°C 时，对聚半乳糖醛酸的最大比活度为 2433.26 U/mg 。这一比活度高于大多数已报道的果胶酸裂解酶。值得注意的是，在 0.4 mM 钙离子存在下，rBvelPL1-Ec 的酶活性提高了 23.28 倍。钙离子对 rBvelPL1-Ec 酶活性的促进作用大于所有已报道的果胶酸裂解酶。用 rBvelPL1-Ec 脱胶后，圈状苎麻纤维的重量减少了 21.27±1.17%，苎麻纤维表面变得更加光滑。此外，经酶解处理后再用 NaOH 处理的圈层苎麻纤维的减重率为 30.47±0.46%。出色的脱胶性能表明，rBvelPL1-Ec 可作为一种有前途的生物催化剂应用于纺织业。

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来源期刊

Enzyme and Microbial Technology 生物-生物工程与应用微生物

CiteScore

7.60

自引率

5.90%

发文量

142

审稿时长

38 days

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.