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designing chimeric enzymes inspired by fungal cellulosomes

Synthetic and Systems Biotechnology 2020年第1期5卷 23-32页

作者： Sean P.Gilmore Stephen P.Lillington Charles H.Haitjema randall de groot Michelle A.O'Malley department of Chemical Engineering University of CaliforniaSanta BarbaraSanta BarbaraCA93106United States MC 5080 University of CaliforniaSanta BarbaraCA93106-5080United States

Cellulosomes are synthesized by anaerobic bacteria and fungi to degrade lignocellulose via synergistic action of multiple enzymes fused to a protein *** templating key protein domains(cohesin and dockerin),designer cellulosomes have been engineered from bacterial motifs to alter the activity,stability,and degradation efficiency of enzyme *** a parts list for fungal cellulosomes from the anaerobic fungi(Neocallimastigomycota)was determined,which revealed sequence divergent fungal cohesin,dockerin,and scaffoldin domains that could be used to expand the available toolbox to synthesize designer *** this work,multi-domain carbohydrate active enzymes(CAZymes)from 3 cellulosome-producing fungi were analyzed to inform the design of chimeric proteins for synthetic cellulosomes inspired by anaerobic *** particular,Piromyces finnis was used as a structural template for chimeric carbohydrate active *** enzymes with retained properties were engineered by combining thermophilic glycosyl hydrolase domains from Thermotoga maritima with dockerin domains from Piromyces *** preserving the protein domain order from ***,chimeric enzymes retained catalytic activity at temperatures over 80°C and were able to associate with cellulosomes purified from anaerobic *** cellulosomes harbor a wide diversity of glycoside hydrolases,each representing templates for the design of chimeric *** conserving dockerin domain position within the primary structure of each protein,the activity of both the catalytic domain and dockerin domain was retained in enzyme *** further,the domain positioning inferred from native fungal cellulosome proteins can be used to engineer multi-domain proteins with non-native favorable properties,such as thermostability.

关键词： Cellulosome Dockerin Scaffoldin Anaerobic fungi Thermophile Enzyme

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