Kayayyaki

Hanyar da tasirin maye gurbi na lokaci guda ya haɗu don canza ayyukan enzymatic ba abu ne mai sauƙi mai iya faɗi ba saboda waɗannan tasirin ba koyaushe suke ƙara ta hanyar layi ba.Don haka, halayen halayen maye gurbi guda ɗaya na ragowar amino acid waɗanda ke ɗaure substrate na iya ba da gudummawa mai mahimmanci ga fahimtar ƙayyadaddun abubuwan enzymes.A cikin beta-glycosidase daga Spodoptera frugiperda (Sfbetagly), duka ragowar Q39 da E451 suna hulɗa tare da substrate kuma wannan yana da mahimmanci don ma'anar ƙayyadaddun kayan aiki.Mutantan Sfbetagly sau biyu (A451E39, S451E39 da S451N39) an shirya su ta hanyar mutagenesis mai jagora, wanda aka bayyana a cikin ƙwayoyin cuta kuma an tsarkake su ta amfani da chromatography na kusanci.Wadannan enzymes an kwatanta su ta hanyar amfani da p-nitrophenyl beta-galactoside da p-nitrophenyl beta-fucoside a matsayin substrates.An yi amfani da rabon k cat/Km don maye gurbi guda ɗaya da biyu na Sfbetagly mai ɗauke da maye gurbi a matsayi a matsayi Q39 da E451 don nuna cewa tasirin makamashi kyauta na ESdouble dagger ( hadaddun yanayin canjin enzyme-transition state complex) na maye gurbi biyu (Gdouble). daggerxy) ba shine jimillar illolin da ake samu daga maye gurbi guda ɗaya ba (Gdouble daggerx da Gdouble daggery).Wannan bambance-bambance a cikin Gdouble dagger yana nuna cewa tasirin maye gurbi guda ɗaya ya zo ɗaya.Don haka, wannan tasirin gama gari yana ƙidaya sau ɗaya kawai a cikin Gdouble daggerxy.Bayanan Crystallographic akan beta-glycosidases sun bayyana kasancewar haɗin haɗin hydrogen bidentate wanda ya haɗa da ragowar Q39 da E451 da ƙungiyar hydroxyl iri ɗaya na substrate.Sabili da haka, duka bayanan thermodynamic da crystallographic suna ba da shawarar cewa ragowar Q39 da E451 suna yin tasiri ga juna akan hulɗar juna tare da ma'auni.