Tan Viet Pham, Truong Chinh Hua, Ngoc An Nguyen, Hanh Thi Dieu Nguyen
{"title":"链霉菌 CNXK100 小型恒温蛋白酶的纯化与特性分析","authors":"Tan Viet Pham, Truong Chinh Hua, Ngoc An Nguyen, Hanh Thi Dieu Nguyen","doi":"10.33073/pjm-2024-014","DOIUrl":null,"url":null,"abstract":"<p><p>Proteases derived from <i>Streptomyces</i> demonstrate numerous commendable properties, rendering it extensively applicable in biotechnology and various industrial sectors. This study focused on the purification and characterization of the thermostable protease obtained from <i>Streptomyces</i> sp. CNXK100. The purified protease exhibited an estimated molecular weight of 27 kDa, with optimal activity at 75°C and pH 8.0. Notably, the enzyme remained active even without any metal ions and fully active in the presence of Na<sup>+</sup>, K<sup>+</sup>, Mg<sup>2+</sup>, and Cu<sup>2+</sup>metal ions. The kinetic parameters were determined with a <i>K<sub>M</sub></i> value of 3.13 mg/ml and a <i>V<sub>max</sub></i> value of 3.28 × 10<sup>6</sup> U/mg. Furthermore, the protease has demonstrated notable stability when subjected to a treatment temperature of up to 65°C for 60 minutes, and across a broad pH range extending from 5.0 to 10.0. This protease also demonstrated resilience against a spectrum of harsh conditions, including exposure to organic solvents, surfactants, bleaching agents, and proteolytic enzymes. Additionally, the enzyme maintained its activity following treatment with commercial detergents, accomplishing complete thrombus lysis at a concentration of 2.50 mg/ml within 4 hours. Remarkably, the protease exhibited stability in terms of activity and protein concentration for 70 days at 4°C. These findings underscore the potential industrial applications of the thermostable protease from <i>Streptomyces</i> sp. CNXK100.</p>","PeriodicalId":94173,"journal":{"name":"Polish journal of microbiology","volume":" ","pages":"155-165"},"PeriodicalIF":0.0000,"publicationDate":"2024-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11192455/pdf/","citationCount":"0","resultStr":"{\"title\":\"Purification and Characterization of a Small Thermostable Protease from <i>Streptomyces</i> sp. CNXK100.\",\"authors\":\"Tan Viet Pham, Truong Chinh Hua, Ngoc An Nguyen, Hanh Thi Dieu Nguyen\",\"doi\":\"10.33073/pjm-2024-014\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Proteases derived from <i>Streptomyces</i> demonstrate numerous commendable properties, rendering it extensively applicable in biotechnology and various industrial sectors. This study focused on the purification and characterization of the thermostable protease obtained from <i>Streptomyces</i> sp. CNXK100. The purified protease exhibited an estimated molecular weight of 27 kDa, with optimal activity at 75°C and pH 8.0. Notably, the enzyme remained active even without any metal ions and fully active in the presence of Na<sup>+</sup>, K<sup>+</sup>, Mg<sup>2+</sup>, and Cu<sup>2+</sup>metal ions. The kinetic parameters were determined with a <i>K<sub>M</sub></i> value of 3.13 mg/ml and a <i>V<sub>max</sub></i> value of 3.28 × 10<sup>6</sup> U/mg. Furthermore, the protease has demonstrated notable stability when subjected to a treatment temperature of up to 65°C for 60 minutes, and across a broad pH range extending from 5.0 to 10.0. This protease also demonstrated resilience against a spectrum of harsh conditions, including exposure to organic solvents, surfactants, bleaching agents, and proteolytic enzymes. Additionally, the enzyme maintained its activity following treatment with commercial detergents, accomplishing complete thrombus lysis at a concentration of 2.50 mg/ml within 4 hours. Remarkably, the protease exhibited stability in terms of activity and protein concentration for 70 days at 4°C. These findings underscore the potential industrial applications of the thermostable protease from <i>Streptomyces</i> sp. CNXK100.</p>\",\"PeriodicalId\":94173,\"journal\":{\"name\":\"Polish journal of microbiology\",\"volume\":\" \",\"pages\":\"155-165\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-04-28\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11192455/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Polish journal of microbiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.33073/pjm-2024-014\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/6/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Polish journal of microbiology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.33073/pjm-2024-014","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/6/1 0:00:00","PubModel":"eCollection","JCR":"","JCRName":"","Score":null,"Total":0}
Purification and Characterization of a Small Thermostable Protease from Streptomyces sp. CNXK100.
Proteases derived from Streptomyces demonstrate numerous commendable properties, rendering it extensively applicable in biotechnology and various industrial sectors. This study focused on the purification and characterization of the thermostable protease obtained from Streptomyces sp. CNXK100. The purified protease exhibited an estimated molecular weight of 27 kDa, with optimal activity at 75°C and pH 8.0. Notably, the enzyme remained active even without any metal ions and fully active in the presence of Na+, K+, Mg2+, and Cu2+metal ions. The kinetic parameters were determined with a KM value of 3.13 mg/ml and a Vmax value of 3.28 × 106 U/mg. Furthermore, the protease has demonstrated notable stability when subjected to a treatment temperature of up to 65°C for 60 minutes, and across a broad pH range extending from 5.0 to 10.0. This protease also demonstrated resilience against a spectrum of harsh conditions, including exposure to organic solvents, surfactants, bleaching agents, and proteolytic enzymes. Additionally, the enzyme maintained its activity following treatment with commercial detergents, accomplishing complete thrombus lysis at a concentration of 2.50 mg/ml within 4 hours. Remarkably, the protease exhibited stability in terms of activity and protein concentration for 70 days at 4°C. These findings underscore the potential industrial applications of the thermostable protease from Streptomyces sp. CNXK100.