链霉菌 CNXK100 小型恒温蛋白酶的纯化与特性分析

Polish journal of microbiology Pub Date : 2024-04-28 eCollection Date: 2024-06-01 DOI:10.33073/pjm-2024-014
Tan Viet Pham, Truong Chinh Hua, Ngoc An Nguyen, Hanh Thi Dieu Nguyen
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摘要

从链霉菌中提取的蛋白酶具有许多值得称道的特性,因此可广泛应用于生物技术和各种工业领域。本研究的重点是纯化和鉴定从链霉菌 CNXK100 中获得的恒温蛋白酶。纯化的蛋白酶分子量约为 27 kDa,在 75°C 和 pH 值为 8.0 时具有最佳活性。值得注意的是,该酶即使在没有任何金属离子的情况下也能保持活性,而在 Na+、K+、Mg2+ 和 Cu2+ 金属离子存在的情况下则完全活跃。经测定,其动力学参数的 KM 值为 3.13 mg/ml,Vmax 值为 3.28 × 106 U/mg 。此外,该蛋白酶在处理温度高达 65°C 的环境中持续 60 分钟,以及在 5.0 至 10.0 的广泛 pH 值范围内都表现出了显著的稳定性。这种蛋白酶还能抵御各种恶劣条件,包括有机溶剂、表面活性剂、漂白剂和蛋白水解酶。此外,这种酶在使用商业洗涤剂处理后仍能保持活性,在 4 小时内以 2.50 毫克/毫升的浓度完全溶解血栓。值得注意的是,这种蛋白酶的活性和蛋白质浓度在 4°C 条件下可稳定 70 天。这些发现强调了来自链霉菌 CNXK100 的恒温蛋白酶的潜在工业应用价值。
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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.

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