Allitol is a valuable drug intermediate and a potential low-calorie sweetener in food industry.However,developing an efficient biotransformation system for large-scale production of allitol remains challenging.Here,a ...Allitol is a valuable drug intermediate and a potential low-calorie sweetener in food industry.However,developing an efficient biotransformation system for large-scale production of allitol remains challenging.Here,a customized multienzyme cascade immobilization system was designed for efficient biocatalytic synthesis of allitol from D-fructose.The system included three immobilized enzymes:amino resin(LX1000)-immobilized Dallulose 3-epimerase(SfDAE)from Sinorhizobium fredii,LX1000-immobilized ribitol dehydrogenase(KpRDH)from Klebsiella pneumoniae,and derived formate dehydrogenase(CbFDH)from Candida boidinii was directional immobilization on bacterial cellulose(BC)employ SpyCatcher/SpyTag modular.By employing a strategy of directional immobilization and SpyCatcher-functionalized BC,the poor cascade cooperativity in the rate-limiting enzyme(CbFDH)was successfully resolved.Under the optimum biotransformation conditions,this system was able to convert 500 g/L of D-fructose into 455 g/L of allitol with conversion yields as high as 90%.Importantly,they also had excellent stability,retaining 60%relative activity after 10 repeated cycles,and preserving 50.2%relative activity after 24 days of storage.Consequently,the design and application of this multienzyme cascade immobilization system successfully provides an effective strategy for the large-scale biosynthesis of allitol.展开更多
基金supported by National Key Research and Develop-ment Program of China(2022YFC2104901)National Natural Science Foundation of China(32372279).
文摘Allitol is a valuable drug intermediate and a potential low-calorie sweetener in food industry.However,developing an efficient biotransformation system for large-scale production of allitol remains challenging.Here,a customized multienzyme cascade immobilization system was designed for efficient biocatalytic synthesis of allitol from D-fructose.The system included three immobilized enzymes:amino resin(LX1000)-immobilized Dallulose 3-epimerase(SfDAE)from Sinorhizobium fredii,LX1000-immobilized ribitol dehydrogenase(KpRDH)from Klebsiella pneumoniae,and derived formate dehydrogenase(CbFDH)from Candida boidinii was directional immobilization on bacterial cellulose(BC)employ SpyCatcher/SpyTag modular.By employing a strategy of directional immobilization and SpyCatcher-functionalized BC,the poor cascade cooperativity in the rate-limiting enzyme(CbFDH)was successfully resolved.Under the optimum biotransformation conditions,this system was able to convert 500 g/L of D-fructose into 455 g/L of allitol with conversion yields as high as 90%.Importantly,they also had excellent stability,retaining 60%relative activity after 10 repeated cycles,and preserving 50.2%relative activity after 24 days of storage.Consequently,the design and application of this multienzyme cascade immobilization system successfully provides an effective strategy for the large-scale biosynthesis of allitol.
