Department of Environmental Science, University of Kashmir, Srinagar, Jammu and Kashmir, India.
Division of Basic Sciences and Humanities, Faculty of Agriculture, SKUAST-Kashmir, Sopore, Jammu and Kashmir, India.
PLoS One. 2023 Mar 30;18(3):e0283677. doi: 10.1371/journal.pone.0283677. eCollection 2023.
In this work, a psychrotrophic bacteria producing cold-active protease, was obtained from Dachigam National Park, an ecologically significant habitat in Western Himalayas owing to its varied endemic and endangered flora and fauna. This isolate was identified as Bacillus sp. HM49 via phenotypic, Gram staining, bio-chemical and 16S rRNA gene identification. Isolate HM49 when tested for proteolytic activity revealed prominent hydrolytic zone with the most production at 20 °C and pH, 8.0 post 72 h incubation. This enzyme was purified, enhancing its specific activity to 61.15 U/mg and its characterization studies revealed it to be a cold-alkaline protease being active in a wide pH (6.0-12) and temperature (5-40 °C) range. Amplification of CAASPR gene of HM49 was performed, followed by enzyme-substrate docking studies and MMGBSA providing details about its type, molecular weight validation as well as functional applications. The purified protease of HM49 was tested for laundry applications and the enzyme was found to be compatible with majority of the detergents tested. Its potential as an eco-friendly detergent additive was further validated by wash performance test as it effectively removed recalcitrant blood stains at a low temperature of 20 °C that could be beneficial for fine garments like silk which preferably need cold washing.
在这项工作中，从达吉甘国家公园获得了一种产生冷活性蛋白酶的嗜冷细菌，由于其多样的特有和濒危动植物群，该公园是西喜马拉雅山具有生态重要意义的栖息地。该分离株通过表型、革兰氏染色、生物化学和 16S rRNA 基因鉴定被鉴定为芽孢杆菌 sp. HM49。分离株 HM49 在测试其蛋白酶活性时，在 20°C 和 pH8.0 下，72 小时孵育后显示出明显的水解区，其产量最高。该酶经过纯化，其比活性提高到 61.15 U/mg，其特性研究表明它是一种冷碱性蛋白酶，在广泛的 pH（6.0-12）和温度（5-40°C）范围内具有活性。对 HM49 的 CAASPR 基因进行了扩增，然后进行酶-底物对接研究和 MMGBSA，提供了有关其类型、分子量验证以及功能应用的详细信息。对 HM49 的纯化蛋白酶进行了洗衣应用测试，结果表明该酶与大多数测试的洗涤剂相容。其作为环保型洗涤剂添加剂的潜力通过洗涤性能测试进一步得到验证，因为它能够有效地去除在 20°C 的低温下顽固的血渍，这对于丝绸等精细衣物可能是有益的，因为丝绸最好需要冷水洗涤。
