{"title":"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。","authors":"Vichi Sicha Irianto, Elif Demirkan, Aynur Aybey Cetinkaya","doi":"10.1080/10826068.2023.2299441","DOIUrl":null,"url":null,"abstract":"<p><p>In this study, the wild-type <i>Bacillus cereus</i> ATA179 was mutagenized by random UV mutagenesis to increase lipase production. The mutant with maximum lipolytic activity was named <i>Bacillus cereus</i> 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, (NH<sub>4</sub>)<sub>2</sub>HPO<sub>4</sub> was the best nitrogen source and CuSO<sub>4</sub> 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. CuSO<sub>4</sub>, CaCl<sub>2</sub>, LiSO<sub>4</sub>, KCl, BaCl<sub>2</sub>, and Tween 20 had an activating effect on the enzyme. <i>V</i><sub>max</sub> and <i>K</i><sub>m</sub> 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.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"918-931"},"PeriodicalIF":2.0000,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"UV mutagenesis for lipase overproduction from <i>Bacillus cereus</i> ATA179, nutritional optimization, characterization and its usability in the detergent industry.\",\"authors\":\"Vichi Sicha Irianto, Elif Demirkan, Aynur Aybey Cetinkaya\",\"doi\":\"10.1080/10826068.2023.2299441\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>In this study, the wild-type <i>Bacillus cereus</i> ATA179 was mutagenized by random UV mutagenesis to increase lipase production. The mutant with maximum lipolytic activity was named <i>Bacillus cereus</i> 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, (NH<sub>4</sub>)<sub>2</sub>HPO<sub>4</sub> was the best nitrogen source and CuSO<sub>4</sub> 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. CuSO<sub>4</sub>, CaCl<sub>2</sub>, LiSO<sub>4</sub>, KCl, BaCl<sub>2</sub>, and Tween 20 had an activating effect on the enzyme. <i>V</i><sub>max</sub> and <i>K</i><sub>m</sub> 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.</p>\",\"PeriodicalId\":20401,\"journal\":{\"name\":\"Preparative Biochemistry & Biotechnology\",\"volume\":\" \",\"pages\":\"918-931\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2024-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Preparative Biochemistry & Biotechnology\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1080/10826068.2023.2299441\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/12/29 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Preparative Biochemistry & Biotechnology","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1080/10826068.2023.2299441","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/12/29 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
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。
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.
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.