Purification and Characterization of a Small Thermostable Protease from Streptomyces sp. CNXK100

Pham, Tan Viet; Hua, Truong Chinh; Nguyen, Ngoc An; Nguyen, Hanh Thi Dieu

doi:10.33073/pjm-2024-014

Abstract

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 K_M value of 3.13 mg/ml and a V_max 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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Purification and Characterization of a Small Thermostable Protease from Streptomyces sp. CNXK100

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