紫外线诱变蜡样芽孢杆菌 ATA179 过量生产脂肪酶、营养优化、特征及其在洗涤剂行业的应用。

IF 2 4区 生物学 Q3 BIOCHEMICAL RESEARCH METHODS Preparative Biochemistry & Biotechnology Pub Date : 2024-08-01 Epub Date: 2023-12-29 DOI:10.1080/10826068.2023.2299441
Vichi Sicha Irianto, Elif Demirkan, Aynur Aybey Cetinkaya
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引用次数: 0

摘要

本研究采用随机紫外诱变的方法对野生型蜡样芽孢杆菌 ATA179 进行诱变,以提高脂肪酶的产量。脂肪酶活性最高的突变体被命名为蜡样芽孢杆菌 EV4。突变菌株(24 h 10.6 U/mL)的产酶量比野生菌株(48 h 6.6 U/mL)高出 60%。研究了营养因素对脂肪酶产量的影响。蔗糖是最佳碳源,(NH4)2HPO4是最佳氮源,CuSO4是最佳金属离子源。突变体 EV4 在改良培养基中的脂肪酶产量增加了 32%。最佳温度和 pH 值分别为 60 °C 和 7.0。CuSO4、CaCl2、LiSO4、KCl、BaCl2 和吐温 20 对酶有激活作用。Vmax 和 Km 值分别为 17.36 U/mL 和 0.036 mM。测定的分子量为 28.2 kDa。脂肪酶的活性在 20 °C 下可稳定 60 天,4 °C 下可稳定 75 天,-20 °C 下可稳定 90 天。研究了脂肪酶在洗涤剂工业中的应用潜力。该酶不受洗涤剂添加剂的影响,但能有效去除被油性物质污染的织物上的污渍。
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UV mutagenesis for lipase overproduction from Bacillus cereus ATA179, nutritional optimization, characterization and its usability in the detergent industry.

In this study, the wild-type Bacillus cereus ATA179 was mutagenized by random UV mutagenesis to increase lipase production. The mutant with maximum lipolytic activity was named Bacillus cereus EV4. The mutant strain (10.6 U/mL at 24 h) produced 60% more enzyme than the wild strain (6.6 U/mL at 48 h). Nutritional factors on lipase production were investigated. Sucrose was the best carbon source, (NH4)2HPO4 was the best nitrogen source and CuSO4 was the best metal ion source. Mutant EV4 showed a 32% increase in lipase production in the modified medium. The optimum temperature and pH were found to be 60 °C and 7.0, respectively. CuSO4, CaCl2, LiSO4, KCl, BaCl2, and Tween 20 had an activating effect on the enzyme. Vmax and Km values were found to be 17.36 U/mL and 0.036 mM, respectively. The molecular weight was determined as 28.2 kDa. The activity of lipase was found to be stable up to 60 days at 20 °C, 75 days at 4 °C, and 90 days at -20 °C. The potential of lipase in the detergent industry was investigated. The enzyme was not affected by detergent additives but was effective in removing stains in fabrics contaminated with oily substances.

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来源期刊
Preparative Biochemistry & Biotechnology
Preparative Biochemistry & Biotechnology 工程技术-生化研究方法
CiteScore
4.90
自引率
3.40%
发文量
98
审稿时长
2 months
期刊介绍: Preparative Biochemistry & Biotechnology is an international forum for rapid dissemination of high quality research results dealing with all aspects of preparative techniques in biochemistry, biotechnology and other life science disciplines.
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