Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia; Biotech ECOZYM Start-up, Business Incubator, Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia; STE JMAL (EJM)-Laundry Detergent Industry, Z.I. Avenue August 13, Z.I. Poudriere 1, P.O. Box 407, Boustene, Sfax 3000, Tunisia.
Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia; Biotech ECOZYM Start-up, Business Incubator, Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
Int J Biol Macromol. 2019 Jan;121:1227-1239. doi: 10.1016/j.ijbiomac.2018.10.139. Epub 2018 Oct 21.
A novel extracellular protease (SAPRH) was hyper-produced (9000 U/mL) from Bacillus safensis RH12, a newly isolated enzyme from a Tunisian offshore oil field. The enzyme was purified to homogeneity, using salt-precipitation, heat-treatment and FPLC anion-exchange chromatography. The purified enzyme was a monomer of molecular mass of ~28 kDa. The NH-terminal 23 amino-acid sequence of SAPRH showed high homology with those of Bacillus-proteases. SAPRH displayed optimal activity at pH 9 and 60 °C. It was strongly inhibited by phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DFP), indicating that it belongs to the serine-proteases family. Moreover, SAPRH was extremely stable at a broad range of temperature and pH retaining 85% of its activity at 50 °C and 75% at pH 11. The enzyme exhibited excellent stability and compatibility with surfactants and commercial detergents, revealing 90% stability with SDS and 100% stability with Class commercial laundry detergent. One of the most distinctive properties is its catalytic efficiency, which is higher than that of Alcalase 2.5 L, typeDX (commercial enzyme) and SAPB from B. pumilus CBS. Interestingly, the results of the wash performance analysis demonstrated considerably good de-staining at 40 °C for 30 min with low supplementation (500 U/mL). Accordingly, such a protease could be considered as a good detergent-additive in detergent industry.
一种新型的细胞外蛋白酶（SAPRH）从突尼斯近海油田新分离的芽孢杆菌 safensis RH12 中大量产生（9000U/mL）。该酶通过盐沉淀、热处理和 FPLC 阴离子交换层析进行纯化至均一性。纯化后的酶为~28kDa 的单体。SAPRH 的 NH 末端 23 个氨基酸序列与芽孢杆菌蛋白酶具有高度同源性。SAPRH 在 pH 9 和 60°C 时表现出最佳活性。它强烈被苯甲基磺酰氟（PMSF）和二碘丙基氟膦酸盐（DFP）抑制，表明它属于丝氨酸蛋白酶家族。此外，SAPRH 在很宽的温度和 pH 范围内非常稳定，在 50°C 时保留 85%的活性，在 pH 11 时保留 75%的活性。该酶在表面活性剂和商业洗涤剂中表现出极好的稳定性和相容性，在 SDS 中的稳定性为 90%，在 Class 商业洗衣洗涤剂中的稳定性为 100%。其最显著的特性之一是其催化效率，高于 Alcalase 2.5L、typeDX（商业酶）和来自 B. pumilus CBS 的 SAPB。有趣的是，洗涤性能分析结果表明，在 40°C 下 30 分钟低补充（500U/mL）时具有相当好的去污效果。因此，这种蛋白酶可以被认为是洗涤剂工业中的一种良好的洗涤剂添加剂。
