Ibicuruzwa

Gusobanukirwa metabolisme ya mikorobe ikenerwa ningirakamaro mugutezimbere no gutezimbere ingirabuzimafatizo zose zishingiye kuri biocatalytic, kuko itegeka gukora neza.Ibi ni ukuri cyane cyane kuri redox biocatalysis aho ingirabuzimafatizo zikora zikoreshwa kubera cofactor / cosubstrate regenerative ubushobozi endogenous muri host.Recombinant Escherichia coli yakoreshejwe mugutanga umusaruro mwinshi wa proline-4-hydroxylase (P4H), dioxydease itera hydroxylation yubusa L-proline muri trans-4-hydroxy-L-proline hamwe na ketoglutarate (a-KG) nka cosubstrate.Muri iyi selile yose ya biocatalysis, metabolisme yo hagati ya karubone itanga cosubstrate isabwa a-KG, igahuza imikorere ya P4H ya biocatalytic yibikorwa bya carbone metabolism hamwe nibikorwa bya metabolike.Mugukoresha ibikoresho byubuzima bwibigeragezo no kubara, nkubwubatsi bwa metabolike na (13) C-metabolic flux analyse ((13) C-MFA), twakoze iperereza kandi tunasobanura muburyo bwa physiologique, metabolic, na bioenergetique igisubizo cya biocatalyst selile yose. kuri bioconversion igamije kandi ikagaragaza inzitizi zishoboka za metabolike kugirango habeho ubundi buryo bwo gukora inzira zifatika.Umuyoboro wa E. coli udafite imbaraga za proline wubatswe no gusiba genA putoding ya proline dehydrogenase.Uturemangingo twose twibinyabuzima hamwe niyi miterere ya mutant ntabwo byatumye habaho hydroxylation ya proline yuzuye gusa ahubwo byanakubye kabiri igipimo cyihariye cya trans-4-L-hydroxyproline (hyp), ugereranije nubwoko bwishyamba.Isesengura ry’imyuka ya karubone binyuze muri metabolisme yo hagati y’imihindagurikire y’imiterere ya mutant ryerekanye ko kwiyongera kwa K-KG ku bikorwa bya P4H bitigeze byongera ingufu za A-KG zitanga umusaruro, ibyo bikaba byerekana imikorere ya TCA igenzurwa cyane mu bihe byakozwe.Mu bwoko bw'ishyamba, synthesis ya P4H na catalizike byatumye igabanuka ry'umusaruro wa biomass.Igishimishije, umurongo wa ΔputA wongeyeho indishyi zijyanye na ATP na NADH mugabanya ingufu zokubungabunga mukigereranyo cyo hasi ya glucose yo gufata, aho kongera ibikorwa bya TCA. PutA knockout muri recombinant E. coli BL21 (DE3) (pLysS) wasangaga kuba ibyiringiro kubisubizo bya P4H bitanga umusaruro gusa mubijyanye numusaruro wa biotransformation gusa, ariko no kubijyanye nigipimo cyibinyabuzima bihindagurika no gufata protine hamwe numusaruro wa hyp ku isoko yingufu.Ibisubizo byerekana ko, kuri putA knockout, guhuza TCA-cycle kugirango bigabanye hydroxylation ikoresheje cosubstrate a-KG ihinduka ikintu cyingenzi kibuza kandi intego yo kurushaho kunoza imikorere ya biotransformations zishingiye kuri KG.