耐盐链霉菌(Streptomyces sp. VITGS3)碱性蛋白酶的工艺优化、提取及其在隐形眼镜清洁剂中的应用

IF 3.5 Q3 Biochemistry, Genetics and Molecular Biology
Gargi Sarkar, K. Prem Anand, M.A. Jayasri, K. Suthindhiran
{"title":"耐盐链霉菌(Streptomyces sp. VITGS3)碱性蛋白酶的工艺优化、提取及其在隐形眼镜清洁剂中的应用","authors":"Gargi Sarkar,&nbsp;K. Prem Anand,&nbsp;M.A. Jayasri,&nbsp;K. Suthindhiran","doi":"10.1016/j.jgeb.2025.100459","DOIUrl":null,"url":null,"abstract":"<div><div>Marine halotolerant actinobacteria are robust microbes poorly explored and barely cultivable in nature. They are a trove of various secondary metabolites and enzymes, especially the alkaline proteases withstanding higher temperatures, pH, and salinity, making them an ideal source with versatile commercial and therapeutic values. This study focuses on extracting and optimizing alkaline protease production from <em>Streptomyces</em> sp. VITGS3 isolated from Puthuvypeen, Kerala. The protease production was optimized by Response Surface Methodology (RSM) using the Box-Behnken model, which used rice bran, wheat bran, skim milk, and casein as substrates. The maximum protease was produced (357 U/mL) using wheat bran (5.5 % w/v) as substrate at pH 9 and incubated at 45 °C for 9 days. The Michaelis-Menten model’s enzyme kinetics exhibited a K<sub>m</sub> value of 1.42 µM, a V<sub>max</sub> of 201.64 µM·min<sup>−1</sup>, V<sub>0</sub> of 5.59 µM·min<sup>−1</sup>, and K<sub>cat</sub> 70013.89 min<sup>−1</sup> suggesting a higher affinity of the enzyme for the substrate (1 % w/v casein). In addition, the protease was inhibited by the phenylmethylsulphonyl fluoride (PMSF), suggesting it belongs to the serine protease family. Finally, the application studies as contact lens cleaners showcased that the isolated protease effectively degraded the protein deposits present in the artificial tear solution without affecting the light transmittance. This is a milestone in the implication of protease on therapeutic applications and further studies on protein specificity, sustained releases, and combination strategies, resulting in crucial challenges in long-term studies, cross-reactivity, storage, and cost-effectiveness.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 1","pages":"Article 100459"},"PeriodicalIF":3.5000,"publicationDate":"2025-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Process optimization and extraction of alkaline protease from halotolerant Streptomyces sp. VITGS3 and its use as a contact lens cleaner\",\"authors\":\"Gargi Sarkar,&nbsp;K. Prem Anand,&nbsp;M.A. Jayasri,&nbsp;K. Suthindhiran\",\"doi\":\"10.1016/j.jgeb.2025.100459\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>Marine halotolerant actinobacteria are robust microbes poorly explored and barely cultivable in nature. They are a trove of various secondary metabolites and enzymes, especially the alkaline proteases withstanding higher temperatures, pH, and salinity, making them an ideal source with versatile commercial and therapeutic values. This study focuses on extracting and optimizing alkaline protease production from <em>Streptomyces</em> sp. VITGS3 isolated from Puthuvypeen, Kerala. The protease production was optimized by Response Surface Methodology (RSM) using the Box-Behnken model, which used rice bran, wheat bran, skim milk, and casein as substrates. The maximum protease was produced (357 U/mL) using wheat bran (5.5 % w/v) as substrate at pH 9 and incubated at 45 °C for 9 days. The Michaelis-Menten model’s enzyme kinetics exhibited a K<sub>m</sub> value of 1.42 µM, a V<sub>max</sub> of 201.64 µM·min<sup>−1</sup>, V<sub>0</sub> of 5.59 µM·min<sup>−1</sup>, and K<sub>cat</sub> 70013.89 min<sup>−1</sup> suggesting a higher affinity of the enzyme for the substrate (1 % w/v casein). In addition, the protease was inhibited by the phenylmethylsulphonyl fluoride (PMSF), suggesting it belongs to the serine protease family. Finally, the application studies as contact lens cleaners showcased that the isolated protease effectively degraded the protein deposits present in the artificial tear solution without affecting the light transmittance. This is a milestone in the implication of protease on therapeutic applications and further studies on protein specificity, sustained releases, and combination strategies, resulting in crucial challenges in long-term studies, cross-reactivity, storage, and cost-effectiveness.</div></div>\",\"PeriodicalId\":53463,\"journal\":{\"name\":\"Journal of Genetic Engineering and Biotechnology\",\"volume\":\"23 1\",\"pages\":\"Article 100459\"},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2025-01-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Genetic Engineering and Biotechnology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1687157X25000034\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Genetic Engineering and Biotechnology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1687157X25000034","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0

摘要

海洋耐盐放线菌是一种强大的微生物,在自然界中很少被探索和培养。它们是各种次生代谢物和酶的宝库,特别是耐高温、pH值和盐度的碱性蛋白酶,使它们成为具有多种商业和治疗价值的理想来源。研究了产自喀拉拉邦Puthuvypeen的Streptomyces sp. VITGS3的碱性蛋白酶的提取及产酶优化。以米糠、麦麸、脱脂牛奶和酪蛋白为底物,采用Box-Behnken模型,采用响应面法(RSM)优化蛋白酶产量。以麦麸(5.5% w/v)为底物,pH为9,45°C孵育9 d,产生最大蛋白酶(357 U/mL)。Michaelis-Menten模型的酶动力学Km值为1.42µM, Vmax为201.64µM·min - 1, V0为5.59µM·min - 1, Kcat 70013.89 min - 1表明该酶对底物(1% w/v酪蛋白)具有较高的亲和力。此外,该蛋白酶被苯基甲基磺酰氟(PMSF)抑制,表明它属于丝氨酸蛋白酶家族。最后,作为隐形眼镜清洁剂的应用研究表明,分离的蛋白酶在不影响透光率的情况下有效地降解了人工泪液中存在的蛋白质沉积物。这是蛋白酶在治疗应用和蛋白质特异性、持续释放和联合策略的进一步研究中的一个里程碑,在长期研究、交叉反应性、储存和成本效益方面带来了重大挑战。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Process optimization and extraction of alkaline protease from halotolerant Streptomyces sp. VITGS3 and its use as a contact lens cleaner

Process optimization and extraction of alkaline protease from halotolerant Streptomyces sp. VITGS3 and its use as a contact lens cleaner
Marine halotolerant actinobacteria are robust microbes poorly explored and barely cultivable in nature. They are a trove of various secondary metabolites and enzymes, especially the alkaline proteases withstanding higher temperatures, pH, and salinity, making them an ideal source with versatile commercial and therapeutic values. This study focuses on extracting and optimizing alkaline protease production from Streptomyces sp. VITGS3 isolated from Puthuvypeen, Kerala. The protease production was optimized by Response Surface Methodology (RSM) using the Box-Behnken model, which used rice bran, wheat bran, skim milk, and casein as substrates. The maximum protease was produced (357 U/mL) using wheat bran (5.5 % w/v) as substrate at pH 9 and incubated at 45 °C for 9 days. The Michaelis-Menten model’s enzyme kinetics exhibited a Km value of 1.42 µM, a Vmax of 201.64 µM·min−1, V0 of 5.59 µM·min−1, and Kcat 70013.89 min−1 suggesting a higher affinity of the enzyme for the substrate (1 % w/v casein). In addition, the protease was inhibited by the phenylmethylsulphonyl fluoride (PMSF), suggesting it belongs to the serine protease family. Finally, the application studies as contact lens cleaners showcased that the isolated protease effectively degraded the protein deposits present in the artificial tear solution without affecting the light transmittance. This is a milestone in the implication of protease on therapeutic applications and further studies on protein specificity, sustained releases, and combination strategies, resulting in crucial challenges in long-term studies, cross-reactivity, storage, and cost-effectiveness.
求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Journal of Genetic Engineering and Biotechnology
Journal of Genetic Engineering and Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
5.70
自引率
5.70%
发文量
159
审稿时长
16 weeks
期刊介绍: Journal of genetic engineering and biotechnology is devoted to rapid publication of full-length research papers that leads to significant contribution in advancing knowledge in genetic engineering and biotechnology and provide novel perspectives in this research area. JGEB includes all major themes related to genetic engineering and recombinant DNA. The area of interest of JGEB includes but not restricted to: •Plant genetics •Animal genetics •Bacterial enzymes •Agricultural Biotechnology, •Biochemistry, •Biophysics, •Bioinformatics, •Environmental Biotechnology, •Industrial Biotechnology, •Microbial biotechnology, •Medical Biotechnology, •Bioenergy, Biosafety, •Biosecurity, •Bioethics, •GMOS, •Genomic, •Proteomic JGEB accepts
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信