从蜡样芽孢杆菌(Bacillus cereusATCC 14579)中克隆、纯化和鉴定重组戊二酸酶的功能,以改善洗涤剂性能

IF 1.6 4区 工程技术 Q3 CHEMISTRY, APPLIED
Asma Zafar, Ammara Masood, Ziaur Rahman, Attia Hamid, Mah Hoor Javed, Amna Zulfiqar, Samreen Fatima, Madood Makhdoom, Muhammad Nauman Aftab
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引用次数: 0

摘要

本研究概述了利用 pET-25b (+) 表达载体将蜡样芽孢杆菌 ATCC 14579 的重组戊二酸酶克隆、表达和鉴定到大肠杆菌 BL21(DE3) 中的方法。利用硫酸铵沉淀法和固定金属离子亲和层析法(IMAC)纯化重组的葡聚糖酶。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)测定,纯化后的葡聚糖酶的分子量为 95 kDa。纯化的重组戊二酸酶具有显著的热稳定性,在高达 90°C 的温度下 4 小时仍能保持其结构完整性和功能性。相反,Co2+ 和 EDTA(乙二胺四乙酸)的存在会抑制酶的活性。纯化的纤维素酶在有机溶剂中表现出显著的耐受性。在 SDS(十二烷基硫酸钠)存在的情况下,酶的活性明显降低,而 β-巯基乙醇和吐温-60 并没有对酶的活性和稳定性产生重大影响,这表明它有可能应用于洗涤剂领域。这一发现为提高市场上现有洗涤剂的功效提供了一种潜在的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cloning, purification, and functional characterization of recombinant pullulanase from Bacillus cereusATCC 14579 for improved detergent performance
The present study outlines the approach that was employed for cloning, expression, and characterization of the recombinant pullulanase enzyme from Bacillus cereus ATCC 14579 into Escherichia coli BL21(DE3) using pET‐25b (+) expression vector. The recombinant pullulanase enzyme was purified using ammonium sulfate precipitation and immobilized metal ion affinity chromatography (IMAC). The molecular mass of the purified pullulanase enzyme was measured using sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) as 95 kDa. The purified recombinant pullulanase enzyme demonstrated significant thermal stability, maintaining its structural integrity and functionality at temperatures as high as 90°C over a period of 4 h. The inclusion of divalent metal ions, specifically Ca2+ and Mg2+, had a positive effect on the activity of the pullulanase enzyme. Conversely, the presence of Co2+ and EDTA (Ethylene Diamine Tetra Acetic acid) resulted in suppression of the enzyme activity. The purified pullulanase enzyme demonstrated remarkable resistance when exposed to organic solvents. The enzyme activity was notably decreased in the presence of SDS (Sodium Dodecyl Sulfate) while β‐mercaptoethanol and tween‐60 did not substantially affect the enzyme activity and stability which suggest its potential applicability in the detergent sector. This discovery indicates a potential approach to improve the effectiveness of currently available detergents in the marketplace.
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来源期刊
Journal of Surfactants and Detergents
Journal of Surfactants and Detergents 工程技术-工程:化工
CiteScore
3.80
自引率
6.20%
发文量
68
审稿时长
4 months
期刊介绍: Journal of Surfactants and Detergents, a journal of the American Oil Chemists’ Society (AOCS) publishes scientific contributions in the surfactants and detergents area. This includes the basic and applied science of petrochemical and oleochemical surfactants, the development and performance of surfactants in all applications, as well as the development and manufacture of detergent ingredients and their formulation into finished products.
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