摘要
为了提高猪胰脂肪酶(PPL)的活性和稳定性,采用一系列具有不同长度疏水烷基的脯氨酸离子液体对PPL进行了修饰。结果表明,所有修饰酶的各项酶学性能都得到了提高,综合来说,[ProC_(8)][H_(2)PO_(4)]-PPL表现出较好的催化性能,催化橄榄油水解最适活性达到原酶的2.7倍,在50℃保存2 h后的剩余酶活是原酶的5.0倍,催化效率K_(cat)·K_(m)^(-1)是原酶的4.4倍,催化α-苯乙醇转酯化的对映体选择性是原酶的1.6倍。光谱学表征揭示了PPL荧光基团微环境及二级结构的变化,这些变化与修饰酶的活性和稳定性的提高有关。与以往报道的结果相比,新型脯氨酸功能离子液体对脂肪酶催化性能的改善更为有效。
A series of proline ionic liquids possessing hydrophobic alkyl of different lengths were used to modify porcine pancreatic lipase(PPL)for improving its activity and stability.The results indicated that the enzymatic properties of all modified lipases were enhanced and[ProC_(8)][H_(2)PO_(4)]-PPL showed the best catalytic performance.The optimal catalytic activity of hydrolyzing olive oil was 2.7 times compared with the native lipase,and the residual enzyme activity was 5.0 times after being stored at 50℃ for 2 h.The catalytic efficiency K_(cat)·K_(m)^(-1)was 4.4 times that of the native lipase,and the enantioselectivity ofα-phenylethanol esterification increased by 1.6 times.The spectroscopic characterization revealed the changes of secondary structure and fluorophore microenvironment of PPL,which were associated with the increase of activity and stability of the modified enzyme.Compared with results previously reported,the novel proline ionic liquid provides a more effective improvement in the catalytic performance of lipase.
作者
张晓光
裴新华
伍慧兰
鲁泽平
胡燚
ZHANG Xiao-guang;PEI Xin-hua;WU Hui-lan;LU Ze-ping;HU Yi(State Key Laboratory of Materials-Oriented Chemical Engineering,School of Pharmaceutical,Nanjing Tech University,Nanjing 211816,China)
出处
《高校化学工程学报》
EI
CAS
CSCD
北大核心
2023年第2期268-275,共8页
Journal of Chemical Engineering of Chinese Universities
基金
国家自然科学基金(22178174,21676143)
江苏省先进生物制造协同创新项目(XTC2206)。
关键词
化学修饰
猪胰脂肪酶
离子液体
稳定性
chemical modification
porcine pancreatic lipase
ionic liquid,stability
