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Maitiro ekusangana nawo mhedzisiro yekuchinja panguva imwe chete kuchinja enzymatic chiitiko haataurike zviri nyore nekuti izvi zvinokonzeresa hazviwanzo kuwedzera nenzira yakatsetseka.Nokudaro, kufananidzwa kwemigumisiro yekuchinja panguva imwe chete yeamino acid residues inosunga substrate inogona kuita zvakakosha pakunzwisisa kwe substrate chaiyo ye enzymes.Mu beta-glycosidase kubva kuSpodoptera frugiperda (Sfbetagly), zvose zvakasara Q39 uye E451 zvinodyidzana ne substrate uye izvi zvakakosha pakutsanangura substrate chaiyo.Kaviri shanduko yeSfbetagly (A451E39, S451E39 uye S451N39) yakagadzirwa nesaiti-inotungamirwa mutagenesis, inoratidzirwa mubhakitiriya uye yakacheneswa uchishandisa affinity chromatography.Aya ma enzymes aionekwa achishandisa p-nitrophenyl beta-galactoside uye p-nitrophenyl beta-fucoside se substrates.Iyo k katsi/Km reshiyo yeimwe uye yakapetwa kaviri shanduko yeSfbetagly ine saiti-inotungamirwa shanduko panzvimbo Q39 uye E451 yakashandiswa kuratidza kuti mhedzisiro pasimba remahara reESdouble dagger (enzyme-transition state complex) yekaviri shanduko (Gdouble). daggerxy) haisi hwerengedzo yemhedzisiro inokonzerwa nekuchinja kumwe chete (Gdouble daggerx uye Gdouble daggery).Uyu mutsauko muGdouble dagger unoratidza kuti mhedzisiro yekuchinja kumwe kunopindirana zvishoma.Nekudaro, izvi zvinowanzoitika zvinoverengera kamwe chete muGdouble daggerxy.Crystallographic data pabeta-glycosidases inoratidza kuvepo kwebidentate hydrogen bond inosanganisira zvakasara Q39 uye E451 uye yakafanana hydroxyl boka reiyo substrate.Naizvozvo, zvese zviri zviviri thermodynamic uye crystallographic data inoratidza kuti zvisaririra Q39 uye E451 zvinopesvedzera zvakafanana pakudyidzana kwavo kwakasiyana neiyo substrate.