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嗜碱短杆菌来源的低温活性胞外碱性蛋白酶:酶的生产及其工业应用。

Cold-active extracellular alkaline protease from an alkaliphilic Stenotrophomonas maltophilia: production of enzyme and its industrial applications.

机构信息

Protein Research Laboratory, Department of Biotechnology, Integral University, Lucknow 226026, India.

出版信息

Can J Microbiol. 2009 Nov;55(11):1294-301. doi: 10.1139/w09-089.

Abstract

A novel psychro-tolerant bacterium Stenotrophomonas maltophilia (MTCC 7528) with an ability to produce extracellular, cold-active, alkaline, and detergent-stable protease was isolated from soil samples obtained from Gangotri glacier, Western Himalaya, India. The culture conditions for higher protease production were optimized with respect to incubation time, agitation, substrate, pH, and temperature. Maximum protease production of 56.2 U x mL(-1) was achieved in the medium at 20 degrees C and pH 9.0 after 120 h incubation. The protease was partially purified by ion-exchange chromatography and approximately 55-fold purification was achieved. The purified enzyme was a 75 kDa protease with maximum activity and stability at pH 10 and 20 degrees C. The activity of enzyme is stimulated by Mn2+ and inhibited completely by metalloprotease inhibitors, indicating that it is a metalloprotease. The protease showed excellent stability and compatibility with commercial detergents and exhibited high efficiency for the removal of different types of protein-containing stains at low temperature. The wash performance analysis of blood and grass stains on cotton fabric showed an increase in reflectance by 26% and 23%, respectively, after treatment with enzyme in comparison to detergent only. These results indicate that it may be a potential component to use as a detergent additive for cold washing and in environmental bioremediation in cold regions.

摘要

从印度喜马拉雅山西部的冈底斯冰川获得的土壤样本中分离到一种具有产生胞外、耐冷、碱性和去污剂稳定蛋白酶能力的新型嗜冷菌 Stenotrophomonas maltophilia (MTCC 7528)。针对孵育时间、搅拌、底物、pH 值和温度,优化了更高蛋白酶产量的培养条件。在 20°C 和 pH 9.0 的培养基中孵育 120 小时后,可获得最大蛋白酶产量 56.2 U x mL(-1)。蛋白酶通过离子交换色谱法进行部分纯化,获得约 55 倍的纯化。纯化的酶是一种 75 kDa 的蛋白酶,在 pH 10 和 20°C 时具有最大的活性和稳定性。该酶的活性受 Mn2+ 的刺激,被金属蛋白酶抑制剂完全抑制,表明它是一种金属蛋白酶。该蛋白酶在低温下具有出色的稳定性和与商业洗涤剂的兼容性,并对不同类型的含蛋白质污渍具有高效去除能力。棉织物上血渍和草渍的洗涤性能分析表明,与仅使用洗涤剂相比,用酶处理后,反射率分别增加了 26%和 23%。这些结果表明,它可能是一种潜在的洗涤剂添加剂,可用于冷洗和寒冷地区的环境生物修复。

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