Department of Colorectal Surgery, The First Hospital, Jilin University, Changchun 130021, People's Republic of China.
Biotechnol Appl Biochem. 2010 May 10;56(1):1-6. doi: 10.1042/BA20100029.
A novel plasmid (pBSR2) was constructed by incorporating a strong lipase promoter and a terminator into the original pBD64. The lipase gene from Bacillus subtilis strain IFFI10210 was cloned into the plasmid pBSR2 and transformed into B. subtilis A.S.1.1655 to obtain an overexpression strain. The recombinant lipase [BSL2 (B. subtilis lipase 2)] has been expressed from the novel constructed strain and used in kinetic resolution of glycidol through enantioselective transesterification. The effects of reaction conditions on the activity as well as enantioselectivity were investigated. BSL2 showed a satisfying enantioselectivity (E>30) under the optimum conditions [acyl donor: vinyl butyrate; the mole ratio of vinyl butyrate to glycidol was 3:1; organic medium: 1,2-dichloroethane with water activity (a(w))=0.33; temperature 40 degrees C]. The remaining (R)-glycidol with a high enantiomeric purity [ee (enantiomeric excess) >99%] could be obtained when the conversion was approx. 60%. The results clearly show a good potential for industrial application of BSL2 in the resolution of glycidol through enantioselective transesterification.
构建了一种新型质粒 (pBSR2)，通过将强脂肪酶启动子和终止子整合到原始质粒 pBD64 中。将来自枯草芽孢杆菌菌株 IFFI10210 的脂肪酶基因克隆到质粒 pBSR2 中，并转化到枯草芽孢杆菌 A.S.1.1655 中，得到过表达菌株。重组脂肪酶 [BSL2（枯草芽孢杆菌脂肪酶 2）] 已从新型构建菌株中表达，并用于通过对映选择性转酯化动力学拆分缩水甘油。研究了反应条件对活性和对映选择性的影响。在最佳条件下 [酰基供体：丁酸乙烯酯；丁酸乙烯酯与缩水甘油的摩尔比为 3:1；有机介质：水活度 (a(w))=0.33 的 1,2-二氯乙烷；温度 40 摄氏度]，BSL2 表现出令人满意的对映选择性 (E>30)。当转化率约为 60%时，可获得具有高对映体纯度的剩余 (R)-缩水甘油[ee（对映体过量）>99%]。结果清楚地表明，BSL2 在通过对映选择性转酯化拆分缩水甘油方面具有良好的工业应用潜力。
Zotero 插件