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Creation of Bioorthogonal Redox Systems Depending on Nicotinamide Flucytosine Dinucleotide.
MedLine Citation:
PMID:  22098020     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Many enzymes catalyzing biological redox chemistry are depending on the omnipresent cofactor, nicotinamide adenine dinucleotide (NAD). NAD is also involved in various non-redox processes. It remains challenging to disconnect one particular NAD-dependent reaction from all others. Here we present a bioorthogonal system that catalyzes the oxidative decarboxylation of L-malate with a dedicated abiotic cofactor, nicotinamide flucytosine dinucleotide (NFCD). By screening the multi-site saturated mutagenesis libraries of the NAD-dependent malic enzyme (ME), we identified the mutant ME-L310R/Q401C, which showed excellent activity with NFCD, yet marginal activity with NAD. We found that another synthetic cofactor, nicotinamide cytosine dinucleotide (NCD), also displayed similar activity with the ME mutants. Inspired by these observations, we mutated D-lactate dehydrogenase (DLDH) and malate dehydrogenase (MDH) to DLDH-V152R and MDH-L6R, respectively, and both mutants showed fully active with NFCD. When coupled with DLDH-V152R, ME-L310R/Q401C required only a catalytic amount of NFCD to convert L-malate. Our results opened the window to engineer bioorthogonal redox systems for a wide variety of applications in systems biology and synthetic biology.
Authors:
Debin Ji; Lei Wang; Shuhua Hou; Wujun Liu; Jinxia Wang; Qian Wang; Zongbao Kent Zhao
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-11-19
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  -     ISSN:  1520-5126     ISO Abbreviation:  -     Publication Date:  2011 Nov 
Date Detail:
Created Date:  2011-11-21     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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