食品与机械2025，Vol.41Issue(4)：1-7,7.DOI:10.13652/j.spjx.1003.5788.2025.60024
基于分子改造手段提高D-来苏糖异构酶酸性条件异构催化活力
Enhancing the acidic activity of D-lyxose isomerase by molecular modification methods
摘要
Abstract
[Objective]To improve the activity of D-lyxose isomerase under acidic conditions.[Methods]Solvent-accessible surface bioinformatics analysis was performed on surface residues of D-lyxose isomerase derived from Caldanaerobius polysaccharolyticus,and six mutation sites,i.e.,K2D,K2E,K8D,K8E,K18D,and K18E,were designed.Mutants were constructed and subjected to recombinant inducible heterologous expression in Escherichia coli.The recombinant mutant enzymes were isolated and purified using a nickel affinity chromatography column.In vitro comparative experiments were conducted to evaluate the catalytic conversion of D-fructose to D-mannose by the recombinant mutant enzymes at acidic pH 5.5 and near-neutral pH 6.5.[Results]The recombinant mutant enzymes K8D and K8E showed significantly improved catalytic conversion rates of D-fructose,with the conversion rate reaching 140%under optimal conditions(pH 6.5).At pH 5.5,the conversion rate was 1.26 times higher than that of the wild-type enzyme.[Conclusion]Molecular modification successfully enhanced the catalytic activity of D-lyxose isomerase mutants under acidic conditions.关键词
D-甘露糖/D-来苏糖异构酶/分子改造/生物酶法/酸稳定性Key words
D-mannose/D-lyxose isomerase/molecular modification/biological enzyme method/acid stability引用本文复制引用
李巧玲,胡阳,周慧灵,喻勋,文李,吴昊..基于分子改造手段提高D-来苏糖异构酶酸性条件异构催化活力[J].食品与机械,2025,41(4):1-7,7.基金项目
湖南省自然科学基金青年项目(编号:2023JJ40018) （编号:2023JJ40018）
国家自然科学基金青年项目(编号:32201963) （编号:32201963）
湖南省科技创新计划资助(编号:2023RC3137) （编号:2023RC3137）
国家级大学生创新创业训练计划项目(编号:S202310536041) （编号:S202310536041）
