由于广泛的治疗,商业,正在探索来自不同来源的微生物蛋白酶的工业应用。在这方面,分离了黄翅目的肠道微生物群,并检查了蛋白酶的产生。在脱脂乳和明胶琼脂平板上对所有分离物进行了主要和次要筛选。蛋白酶阳性分离株的形态学特征,生物化学,和分子。在20个分离的菌株中,6属于五个不同的属。芽孢杆菌,Priestia,气单胞菌,葡萄球菌,和沙雷氏菌表现出蛋白水解活性。萨瓦芽孢杆菌菌株PRN1表现出最高的蛋白酶产量,因此,脱脂乳琼脂(15±1mm)和明胶(16±1mm)平板中最大的水解透明区。优化后的参数(时间,pH值,温度,碳,氮)的最高蛋白酶活性和微生物生长。safensis菌株PRN1包括72小时(OD600=0.56,1303U/mL),pH8(OD600=0.83,403.29U/mL),40°C(OD600=1.75,1849.11U/mL),果糖(OD600=1.22,1502U/mL),和明胶(OD600=1.88,1015.33U/mL)。使用盐沉淀和凝胶过滤色谱法将酶纯化至均匀。十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)表明,纯化的酶是分子量约33kDa的单体。蛋白酶在pH8和60°C时表现出最佳活性。它被苯甲基磺酰氟(PMSF)强烈抑制,证明它属于丝氨酸蛋白酶家族。酶与表面活性剂和商业洗涤剂的相容性证明了其在洗涤剂工业中的潜在用途。此外,纯化的酶具有抗菌和去除血渍的特性。
Owing to vast therapeutic, commercial, and industrial applications of microbial proteases microorganisms from different sources are being explored. In this regard, the gut microbiota of Monopteruscuchia were isolated and examined for the production of protease. All the isolates were primarily and secondarily screened on skim milk and gelatin agar plates. The protease-positive isolates were characterized morphologically, biochemically, and molecularly. Out of the 20 isolated strains,6 belonging to five different genera viz.Bacillus,Priestia,Aeromonas,Staphylococcus, and Serratia demonstrated proteolytic activity. Bacillussafensis strain PRN1 demonstrated the highest protease production and, thus, the largest hydrolytic clear zones in both skim milk agar (15 ± 1 mm) and gelatin (16 ± 1 mm) plates. The optimized parameters (time, pH, temperature, carbon, nitrogen) for highest protease activity and microbial growth of B.safensis strain PRN1 includes 72 h (OD600 = 0.56,1303 U/mL), pH 8 (OD600 = 0.83, 403.29 U/mL), 40 °C (OD600 = 1.75, 1849.11 U/mL), fructose (OD600 = 1.22, 1502 U/mL), and gelatin (OD600 = 1.88, 1015.33 U/mL). The enzyme was purified to homogeneity using salt-precipitation and gel filtration chromatography. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that the purified enzyme was a monomer of a molecular weight of ∼33 kDa. The protease demonstrated optimal activity at pH 8 and 60 °C. It was strongly inhibited by phenylmethylsulfonyl fluoride (PMSF), demonstrating that it belongs to the serine-proteases family. The compatibility of the enzyme with surfactants and commercial detergents demonstrates its potential use in the detergent industry. Furthermore, the purified enzyme showed antibacterial and blood-stain removal properties.