Gupta Meenu, Choudhury Bijan, Navani Naveen Kumar
Botany Department, J. D. Women's College Patna, Bihar, 800023, India.
Department of Biotechnology, Indian Institute of Technology, Roorkee, Uttarakhand, 247667, India.
Heliyon. 2024 Jan 20;10(3):e25084. doi: 10.1016/j.heliyon.2024.e25084. eCollection 2024 Feb 15.
An unusual haloalkaliphilic bacterium known as sp. strain BNMIITR, which was noticed to produce an extracellular alkaline protease, was found in a soil sample from Northern India's Sambhar Lake. On the generation of protease, the effects of dietary elements including nitrogen and carbon sources, amino acids, and growth conditions like temperature and pH were investigated. When low-cost agricultural by-products were employed as nitrogen sources, the manufacturing of enzymes was significantly boosted. In the present study, protease production was enhanced by 2.94 fold and 2.17 fold. By solvent precipitation and Hydrophobic interaction chromatography (HIC) on Phenyl Sepharose 6 Fast Flow matrix, the enzyme was purified 31.67 fold. It was determined that the apparent molecular mass was 21 kDa. The pH range where the enzyme was most stable was 6.0-12.0, with a temperature of 50 °C as optimum. When there was alkaline earth metals and heavy metals, protease was discovered to be active. It was evident that the enzyme was a serine type of protease because it was active in the presence of a variety of surfactants, oxidizing and reducing chemicals, and phenylmethylsulfonyl fluoride (PMSF) completely inhibited activity. Enzyme exhibited a wide range of substrate specificity. Amazingly, enzyme remained stable both in polar and nonpolar solvents. The most interesting aspect of this enzyme is enhanced activity in polar solvents like dimethylformamide (DMF) and dimethyl sulfoxide (DMSO). It was discovered that the protease was stable and compatible with a number of widely available detergents.
一种名为sp. strain BNMIITR的不寻常嗜碱嗜盐细菌，被发现能产生一种细胞外碱性蛋白酶，它是在印度北部桑巴尔湖的土壤样本中发现的。关于蛋白酶的产生，研究了包括氮源和碳源、氨基酸等营养成分以及温度和pH值等生长条件的影响。当使用低成本农业副产品作为氮源时，酶的产量显著提高。在本研究中，蛋白酶产量提高了2.94倍和2.17倍。通过在苯基琼脂糖6快速流动基质上进行溶剂沉淀和疏水相互作用色谱（HIC），该酶被纯化了31.67倍。测定其表观分子量为21 kDa。该酶最稳定的pH范围是6.0 - 12.0，最适温度为50°C。发现当存在碱土金属和重金属时，蛋白酶具有活性。很明显该酶是丝氨酸型蛋白酶，因为它在多种表面活性剂、氧化和还原化学物质存在下具有活性，并且苯甲基磺酰氟（PMSF）完全抑制其活性。该酶表现出广泛的底物特异性。令人惊讶的是，该酶在极性和非极性溶剂中都保持稳定。这种酶最有趣的方面是在二甲基甲酰胺（DMF）和二甲基亚砜（DMSO）等极性溶剂中活性增强。发现该蛋白酶稳定且与多种常用洗涤剂兼容。
