赖氨酸酰化酶的重组表达及其催化合成ε-月桂酰-L-赖氨酸

来源于链霉菌的赖氨酸酰化酶Sm-ELA能催化赖氨酸和月桂酸在水相中合成月桂酰赖氨酸,避免了采用化学合成法所必需的高温和有机溶剂条件,是一种节能、环境友好的替代方法。构建了过表达链霉菌赖氨酸酰化酶基因的重组质粒pET28a-SmELA和pTrcOmpXK₁₂₂SmELA,分别实现了该酶在大肠杆菌胞内和细胞表面的活性表达。比较两种不同表达方式的效果后,将重组酶应用于催化合成月桂酰赖氨酸的反应中,结果显示,在赖氨酸浓度为50 mmol/L,月桂酸浓度为10 mmol/L时,反应24 h,月桂酸转化率最高达到31.1%。

Recombinant Expression of ε-Lysine Acylase from Streptomyces mobaraensis for Synthesis of Nε-lauroyl-L-lysine

ε-Lysine acylase from Streptomyces mobaraensis(Sm-ELA) can catalyzes hydrolysis of Nε-lauroyl-L-lysine in water with ε-lysine and lauric acid as the substrates. It's an energy-saving and environmentally friendly way that avoids the high temperature and organic solvent by using the chemical method. Two recombinant plasmids which are pET28a-SmELA and pTrcOmpXK₁₂₂SmELA were constructed and a normal expression of intracellular and the cell surface were achieved respectively. The recombinant enzymes were used in catalytic synthesis of Nε-lauroyl-L-lysine and the catalytic efficiency was preliminarily compared. Nε-lauroyl-L-lysine was synthesized from 50 mmol/L ε-lysine and 10 mmol/L lauric acid in an aqueous buffer solution at 37℃. The maximum yield was 31. 1% after 24 h of reaction for 10 mmol/L lauric acid.