Zogulitsa

Momwe zotsatira za masinthidwe anthawi imodzi zimaphatikizidwira kuti zisinthe zochita za enzymatic sizodziwikiratu chifukwa izi sizimawonjezera nthawi zonse motsatira mzere.Chifukwa chake, mawonekedwe a zotsatira za masinthidwe anthawi imodzi a zotsalira za amino acid zomwe zimamangiriza gawo lapansi zitha kuthandizira kwambiri kumvetsetsa gawo la gawo lapansi la michere.Mu beta-glycosidase yochokera ku Spodoptera frugiperda (Sfbetagly), zotsalira zonse za Q39 ndi E451 zimalumikizana ndi gawo lapansi ndipo izi ndizofunikira pakutanthauzira tsatanetsatane wa gawo lapansi.Zosintha ziwiri za Sfbetagly (A451E39, S451E39 ndi S451N39) zidakonzedwa ndi mutagenesis motsogozedwa ndi tsamba, zowonetsedwa ndi mabakiteriya ndikuyeretsedwa pogwiritsa ntchito affinity chromatography.Ma enzymes awa adadziwika pogwiritsa ntchito p-nitrophenyl beta-galactoside ndi p-nitrophenyl beta-fucoside ngati magawo.Chiyerekezo cha k cat/Km cha masinthidwe amodzi ndi awiri a Sfbetagly okhala ndi masinthidwe otsogozedwa ndi tsamba pamalo a Q39 ndi E451 adagwiritsidwa ntchito kuwonetsa kuti mphamvu yaulere ya ESdouble dagger (enzyme-transition state complex) yosinthika kawiri (Gdouble). daggerxy) si kuchuluka kwa zotsatira zobwera chifukwa cha masinthidwe amodzi (Gdouble daggerx ndi Gdouble daggery).Kusiyanaku kwa Gdouble dagger kukuwonetsa kuti zotsatira za masinthidwe amodzi zimadutsana pang'ono.Chifukwa chake, izi zimachitika kamodzi kokha mu Gdouble daggerxy.Zambiri za Crystallographic pa beta-glycosidase zimawulula kukhalapo kwa bidentate hydrogen bond yophatikiza zotsalira Q39 ndi E451 ndi gulu lomwelo la hydroxyl la gawo lapansi.Chifukwa chake, ma data onse a thermodynamic ndi crystallographic akuwonetsa kuti zotsalira za Q39 ndi E451 zimathandizirana pakuyanjana kwawo ndi gawo lapansi.