Khai Minh Technology Group - KMTG, Ho Chi Minh City, Viet Nam; Faculty of Biological Sciences, Nong Lam University, Ho Chi Minh City, Viet Nam.
Research Institute for Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Viet Nam; Faculty of Biological Sciences, Nong Lam University, Ho Chi Minh City, Viet Nam.
Protein Expr Purif. 2024 Mar;215:106419. doi: 10.1016/j.pep.2023.106419. Epub 2023 Dec 16.
A lipase EstA from Bacillus subtilis KM-BS was expressed in Escherichia coli BL21 (DE3) cells. The recombinant enzyme achieved high activity (49.67 U/mL) with protein concentration of 1.29 mg/mL under optimal conditions at the large-scale expression of 6 h and post-induction time at 30 °C using 0.1 mM isopropyl-β-d-thiogalactopyranoside (IPTG). The optimal temperature and pH of the purified enzyme were at 45-55 °C and pH 8.0 - 9.0, respectively. Activity of the purified enzyme was stable in the presence of 1 mM Ca; stimulated by 1 mM Mg and Mn, and inhibited by Fe. A significant amount of fatty acids was released during the hydrolysis of waste cooking oil under the catalysis of purified lipase, indicating that this recombinant lipase showed promise as a suitable candidate in industrial fields, particularly in biodiesel and detergent sector.
枯草芽孢杆菌 KM-BS 的脂肪酶 EstA 在大肠杆菌 BL21（DE3）细胞中表达。在 6 h 的大规模表达和 30°C 的诱导后时间下，使用 0.1 mM 的异丙基-β-D-硫代半乳糖苷（IPTG），在最佳条件下，该重组酶的蛋白浓度为 1.29mg/mL 时，实现了 49.67 U/mL 的高活性。纯化酶的最适温度和 pH 值分别为 45-55°C 和 pH 8.0-9.0。纯化酶在存在 1 mM Ca 时活性稳定，受 1 mM Mg 和 Mn 刺激，受 Fe 抑制。在纯化脂肪酶的催化作用下，废烹调油的水解过程中释放出大量脂肪酸，表明这种重组脂肪酶在工业领域，特别是在生物柴油和洗涤剂领域有很大的应用潜力。
