Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, CAS Key Laboratory of Renewable Energy, Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development, Guangzhou, China.
College of Food Science and Technology, Guangdong Ocean University, Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution, Zhanjiang, China.
Biotechnol Appl Biochem. 2022 Oct;69(5):1812-1820. doi: 10.1002/bab.2247. Epub 2021 Sep 14.
Due to its simple, less by-product and environment friendly properties, enzymatic transesterification of oil with short-chain alcohol to biodiesel, fatty acid methyl esters (FAMEs) is considered to be a promising way of green production and has attracted much attention. In this study, FAMEs were synthesized by an enzymatic method with recombinant lipase as catalysts. A thermophilic Bacillus thermocatenulatus lipase 2 (BTL2) was overexpressed in Escherichia coli BL21(DE3) through relative and quantitative analysis using real-time quantitative PCR. The results suggested that the BTL2 gene was overexpressed in E. coli at the mRNA level, and the recombinant strain harboring a high-copy number vectors was selected and applied to fermentation to produce BTL2 with enzyme activity of 35.54 U/mg cells. The recombinant BTL2 solution exhibited excellent resistance to neutral pH, high temperature, and organic solvents after a certain treatment. Finally, the effects of enzymatic transesterification for preparing biodiesel were studied, using rapeseed oil as raw material, as well as BTL2 solution as catalysts, which resulted in 86.04% yield of FAMEs under 50°C for 36 h. The liquid BTL2 was directly used to prepare FAMEs at a higher temperature efficiently, making the thermophilic BTL2 had the potential application value in biodiesel reproduction subsequently.
由于其简单、副产物少且环境友好的特性，用短链醇酶法对油脂进行酯交换制备生物柴油，即脂肪酸甲酯（FAMEs），被认为是一种很有前途的绿色生产方式，引起了广泛关注。本研究采用酶法，以重组脂肪酶为催化剂合成 FAMEs。通过实时定量 PCR 进行相对和定量分析，在大肠杆菌 BL21(DE3)中过表达了嗜热芽孢杆菌脂肪酶 2（BTL2）。结果表明，BTL2 基因在大肠杆菌中在 mRNA 水平上得到了过表达，选择携带高拷贝数载体的重组菌株进行发酵，以产生酶活为 35.54 U/mg 细胞的 BTL2。经过一定处理后，重组 BTL2 溶液表现出优异的耐中性 pH、高温和有机溶剂的性能。最后，以菜籽油为原料，用 BTL2 溶液作为催化剂，研究了酶法制备生物柴油的效果，在 50°C 下反应 36 h，FAMEs 的产率达到 86.04%。将液态 BTL2 直接用于在较高温度下高效制备 FAMEs，使嗜热 BTL2 随后在生物柴油生产中具有潜在的应用价值。
