食品与发酵工业2023，Vol.49Issue(24)：29-36,8.DOI:10.13995/j.cnki.11-1802/ts.035083
共识序列改造提高南极假丝酵母脂肪酶B的热稳定性
Improving the thermal stability of Candida antarctica lipase B based on consensus sequence modification
摘要
Abstract
Candida antarctica lipase B(CALB)is an enzyme with excellent catalytic efficiency for both water-soluble and non-water-soluble substrates.As a result,it is widely used in various industries such as the synthesis of biodiesel,enantiomeric ketoprofen,and glycerol carbonate,etc.To improve the thermal stability,CALB was aligned with 48 lipase sequences from diverse sources to generate a consensus sequence,and 14 mutations were initially detected that considerably strengthened the conservation of CALB.Subsequently,the combined mutant G114A/A284N(CALBm)with significantly improved thermal stability was determined by site-directed mutation analy-sis.The results showed that the half-life(t1/2)of CALBm at 50 ℃ was increased by 74.7％compared with CALB,reaching 14.5 min;however,the specific activity(16.1 U/mg)and kcat/Km[5 932.3 mmol/(L·min)]decreased by 36.3％and 23％,respectively.Ac-cording to molecular dynamics simulation,the improved hydrophobic interaction(G114 vs M83,G114 vs V84,G114 vs I121)and hydro-gen bonding(A284 vs G41,A284 vs A281)enhanced the overall rigidity and stability of CALBm.Meanwhile,molecular docking analy-sis showed that the decrease in the interaction between CALBm and substrate molecules was the main reason for the reduced activity.These results will benefit the application of CALB at high temperatures.关键词
脂肪酶B/共识序列/热稳定性/酶学性质/分子动力学模拟/分子对接Key words
lipase B/consensus sequence/thermal stability/enzymatic properties/molecular dynamics simulation/molecular docking引用本文复制引用
王亚婵,李阳阳,刘松..共识序列改造提高南极假丝酵母脂肪酶B的热稳定性[J].食品与发酵工业,2023,49(24):29-36,8.基金项目
国家重点研发计划项目(2021YFC2101400) （2021YFC2101400）
