Zymomonas mobilis Ndh kofaktoru specifiskuma mainīšana izmantojot sait-specifisko mutaģenēzi

6533b832fe1ef96bd129a80c

RESEARCH PRODUCT

Zymomonas mobilis Ndh kofaktoru specifiskuma mainīšana izmantojot sait-specifisko mutaģenēzi

Elīna Balodīte

subject

description

Lai pētītu Zymomonas mobilis NAD(P)H dehidrogenāzes substrātu specifiskumu un aktivitāti, izveidots plazmīdas konstrukts ar ndh gēnu. Izmantojot sait-specifisko mutaģenēzi iegūts ndh gēna mutantais variants. Tā produktā (Ndh) 220 pozīcijā Glu nomainīta uz Gln. Veikta Zm6-ndh komplementācija ar natīvo ndh gēnu un tā mutanto variantu. Izmantojot membrānu vezikulas, mērīta NAD(P)H dehidrogenāzes aktivitāte, par elektronu donoriem izmantojot NADH vai NADPH. Mutantās Ndh kinētikas analīze parādīja, ka šim enzīmam ir samazināta Km vērtība par substrātu izmantojot NADPH, pieaugusi attiecība VmaxNADPH/VmaxNADH ,bet absolūtā aktivitāte uz abiem kofaktoriem ir samazinājusies. Audzēšanas eksperimentos parādīts, ka Ndh aktivitāte negatīvi korelē ar augšanas ātrumu. Maģistra darbs izstrādāts Latvijas Universitātes Mikrobioloģijas un Biotehnoloģijas institūtā. Atslēgas vārdi: NAD(P)H dehidrogenāze, Zymomonas mobilis, elpošanas ķēde, sait-specifiskā mutaģenēze

year	journal	country	edition	language
2014-01-01

https://dspace.lu.lv/dspace/handle/7/26911

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