Lihlahisoa

Mokhoa oo litlamorao tsa liphetoho tsa nako e le 'ngoe li kopanang ho fetola tšebetso ea enzymatic ha o bonolo ho tseba esale pele hobane litlamorao tsena ha se kamehla li tlatselletsang ka mokhoa o ikhethileng.Ka hona, sebopeho sa litlamorao tsa liphetoho tsa nako e le 'ngoe tsa masala a amino acid a tlamang karoloana e ka kenya letsoho haholo kutloisisong ea likarolo tse ikhethang tsa li-enzyme.Ho beta-glycosidase e tsoang ho Spodoptera frugiperda (Sfbetagly), masala ka bobeli a Q39 le E451 a sebelisana le substrate mme sena se bohlokoa bakeng sa ho hlalosa boleng ba substrate.Liphetoho tse habeli tsa Sfbetagly (A451E39, S451E39 le S451N39) li lokisitsoe ke mutagenesis e lebisitsoeng sebakeng, e hlahisoang ka baktheria 'me ea hloekisoa ka ho sebelisa affinity chromatography.Li-enzyme tsena li ne li khetholloa ka ho sebelisa p-nitrophenyl beta-galactoside le p-nitrophenyl beta-fucoside e le likaroloana.Karo-karolelano ea k cat/Km bakeng sa phetoho e le 'ngoe le e habeli ea Sfbetagly e nang le liphetoho tse lebisitsoeng sebakeng sa Q39 le E451 e sebelisitsoe ho bonts'a hore phello ea matla a mahala a ESdouble dagger (enzyme-transition state complex) ea liphetoho tse habeli (Gdouble). daggerxy) ha se kakaretso ea litlamorao tse hlahisoang ke phetoho e le 'ngoe (Gdouble daggerx le Gdouble daggery).Phapang ena ho Gdouble dagger e bonts'a hore litlamorao tsa phetoho e le 'ngoe li kopana hanyane.Kahoo, phello ena e tloaelehileng e baloa hang feela ho Gdouble daggerxy.Lintlha tsa Crystallographic mabapi le beta-glycosidase li senola boteng ba bond ea hydrogen ea bidentate e kenyelletsang masala Q39 le E451 le sehlopha se tšoanang sa hydroxyl sa substrate.Ka hona, lintlha tse peli tsa thermodynamic le crystallographic li fana ka maikutlo a hore masala a Q39 le E451 a na le tšusumetso e tšoanang litšebelisanong tsa bona tse fapaneng le substrate.