Department of Molecular Biology and Genetics, Faculty of Science, Erzurum Technical University, Erzurum, Turkey.
Arch Microbiol. 2020 Jan;202(1):153-159. doi: 10.1007/s00203-019-01728-7. Epub 2019 Sep 20.
In this study, it was aimed to determine the ability to produce protease enzyme of Thermomonas haemolytica isolated from geothermal Nenehatun hot spring in Turkey and utilization of this enzyme in the detergent industry to remove protein stains. The protease-producing strains were screened from hot springs, and a potential strain was identified as T. haemolytica according to morphological, physiological and biochemical characteristics and sequence of 16S rRNA gene. Maximum protease activity was observed at 55 °C and pH 9.0 at 72 h of incubation. Activity was very stable between 50 and 65 °C and pH 8.0-10.0, respectively. The enzyme activity was significantly inhibited by PMSF and partly inhibited by EDTA, EGTA, SDS, and urea. Some divalent metal ions such as Ca, Mg, and Mn increased the enzyme activity, while Zn and Cu decreased. Michaelis-Menten constant (K) and maximum velocity (V) values were calculated by Lineweaver-Burk plot as 125 EU/ml and 1262 mg/ml, respectively. The biochemical characterization of the protease obtained from T. haemolytica was performed and applied on the blood and grass-stained fabrics with detergent to evaluate the stain removal performance of the enzyme. It was observed that the application of detergent with enzyme was more effective than the detergent without enzyme to clean up the stained fabrics. This is the first report of characterization of the protease of T. haemolytica. According to results obtained from this study, this new strain is a promising candidate for industrial applications in production of detergent.
在这项研究中，旨在确定从土耳其内内哈顿温泉分离的嗜热单胞菌产生蛋白酶酶的能力，并将该酶应用于去污剂行业以去除蛋白质污渍。从温泉中筛选出产生蛋白酶的菌株，并根据形态学、生理学和生物化学特征以及 16S rRNA 基因序列，将潜在的菌株鉴定为嗜热单胞菌。在 55°C 和 pH9.0 下孵育 72 小时时观察到最大蛋白酶活性。在 50 至 65°C 和 pH8.0-10.0 之间，活性非常稳定。该酶的活性被 PMSF 显著抑制，被 EDTA、EGTA、SDS 和尿素部分抑制。一些二价金属离子如 Ca、Mg 和 Mn 增加了酶的活性，而 Zn 和 Cu 则降低了酶的活性。通过 Lineweaver-Burk 作图计算米氏常数（K）和最大速度（V）值分别为 125 EU/ml 和 1262 mg/ml。对从嗜热单胞菌获得的蛋白酶进行了生化特性分析，并应用于含有去污剂的血液和草渍织物上，以评估酶的去污性能。观察到，与不含酶的去污剂相比，含有酶的去污剂更有效地清洁染色织物。这是首次报道嗜热单胞菌蛋白酶的特性。根据本研究的结果，该新菌株是在生产去污剂中工业应用的有前途的候选菌株。
