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1、REVIEWpubs.acs.org/acscatalysisEngineeringofBiocatalysts:fromEvolutiontoCreationMaureenB.QuinandClaudiaSchmidt-Dannert*DepartmentofBiochemistry,MolecularBiology&Biophysics,UniversityofMinnesota,140GortnerLaboratory,1479GortnerAvenue,SaintPaul,Minnesota55108,UnitedStatesABSTRACT:Enzymesareincreasin
2、glybeingusedinanindustrialsettingasacheapandenvironmentallyfriendlyalternativetochemicalcatalysts.Toproducetheidealbiocatalyst,naturalenzymesoftenrequireoptimizationtoincreasetheircatalyticefficienciesandspecificitiesunderaparticularrangeofreactionconditions.Anumberofenzymeengineeringstrategiesarecur
3、rentlyemployedtomodifybiocatalysts,improvingtheirsuitabilityforlarge-scaleindustrialapplications.Theseincludevariousdirectedevolutiontechniques,semirationaldesigntechniques,andmorerecently,thedenovodesignofnovelenzymes.Advancesinmutantlibrarydesign,high-throughputselectionprocesses,andtheintroduct
4、ionofpowerfulcomputeralgorithmshaveallcontributedtothecurrentexponentialgrowthofthefieldofenzymeengineering.Thisreviewarticleaimstopresentsomeofthecurrentlyemployedstrategiesforenzymeengineeringandattemptstohighlightthemostrecentadvancesinmethodology.KEYWORDS:biocatalysts,enzymeengineering,directed
5、evolution,semirationalde-sign,denovodesign’INTRODUCTIONexamplesofsuccessfulengineeringofbiocatalystspresented,andrecentadvancesinthefieldwillalsobehighlighted.Enzymesareexcellentcatalysts.Millionsofyearsofevolutionhasaffordedenzymeswiththeabilitytoacceleratetherateofchemicalreactionswithahighdegreeo
6、fefficiency,selectivity,’DIRECTEDEVOLUTIONandspecificity.Assuch,theuseofenzymesasbiocatalystsforDirectedevolutionisbasedupontheprincipleofnaturalindustrialprocessesisaneconomicallyandenvironmentallyattr-evolution,wherebytheincorporationofrandommutationsinto1activeoption.However,naturalenzymesareoften
7、unabletothesequenceofanenzymeallowsthecreationalargemutantfunctioninthenon-naturalconditionsdictatedbymanyindus-library(103106mutants)displayingahighlevelofsequencetrialreactions,whichfrequentlyrequireelevatedte
