Ibicuruzwa

Uburyo ingaruka ziterwa na mutation icyarimwe zihuza kugirango zihindure ibikorwa byimisemburo ntabwo byoroshye guhanurwa kuko izi ngaruka ntabwo buri gihe ziyongera muburyo bumwe.Kubwibyo, kuranga ingaruka ziterwa no guhinduka icyarimwe ibisigisigi bya aminide acide ihuza substrate birashobora gutanga umusanzu wingenzi mugusobanukirwa umwihariko wa substrate yihariye.Muri beta-glycosidase yo muri Spodoptera frugiperda (Sfbetagly), ibisigisigi byombi Q39 na E451 bikorana na substrate kandi ibi nibyingenzi mugusobanura umwihariko wa substrate.Impinduka ebyiri za Sfbetagly (A451E39, S451E39 na S451N39) zateguwe na mutagenezi uyobowe na site, zigaragara muri bagiteri kandi zezwa hakoreshejwe chromatografiya.Iyi misemburo yaranzwe no gukoresha p-nitrophenyl beta-galactoside na p-nitrophenyl beta-fucoside nka substrate.Ikigereranyo cya k injangwe / Km kuri mutant imwe na kabiri ya Sfbetagly ikubiyemo ihinduka ryerekanwe kurubuga kumwanya wa Q39 na E451 yakoreshejwe kugirango yerekane ko ingaruka zingufu zubusa za ESdouble hanjeri (enzyme-inzibacyuho ya leta) ya mutation ebyiri (Gdouble Daggerxy) ntabwo ari igiteranyo cyingaruka ziterwa na mutation imwe (Gdouble daggerx na Gdouble daggery).Itandukaniro muri Gdouble imipanga yerekana ko ingaruka za mutation imwe zuzuzanya igice.Kubwibyo, iyi ngaruka rusange ibarwa rimwe gusa muri Gdouble daggerxy.Amakuru ya Crystallographic kuri beta-glycosidase yerekana ko hariho hydrogène ya bidentate irimo ibisigisigi Q39 na E451 hamwe nitsinda rimwe rya hydroxyl ya substrate.Kubwibyo, byombi bya termodinamike hamwe na kristu yerekana ko ibisigisigi Q39 na E451 bigira uruhare runini mubikorwa byabo hamwe na substrate.