DICP OpenIR
Subject Area生物工程
Assembling of Bioorthogonal Redox Systems Depending on an NAD Analog
Zhao ZB(赵宗保); Ji DB(纪德彬); Wang L(王磊); Liu WJ(刘武军); Wang JX(王金霞); Hou SH(侯淑华); Wang Q(王倩)
Source PublicationBITS 2nd Symposium on Enzymes and Biocatalysis-2011
Conference NameBITS 2nd Symposium on Enzymes & Biocatalysis-2011
Conference Date2011-4-25
2011
Conference Place大连
Pages61-0
Publisher待补充
Publication Place待补充
Cooperation Status分会口头报告
Department1816
Funding Organization中国医药生物技术协会
AbstractBiological redox chemistry is catalyzed by numerous enzymes depending largely on a few cofactors. Nicotinamide adenine dinucleotide (NAD) is one of the most important cofactors, is also involved in diverse non-redox processes. It remains challenging to disconnect one redox reaction from the other NAD-dependent cellular events. 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 multi-site saturated mutagenesis libraries of the NAD-dependent malic enzyme (ME) from Escherichia coli, we identified the mutant ME-L3I0R/Q401C, activity of which was excellent with NFCD, but marginal with NAD. Moreover, wild type ME showed little activity with NFCD. Based on the insights abstracted from this process, we were able to generate a mutant of D-lactate dehydrogenase (DLDH) from Lacrobacillus helvettcus, DLDH-V152R, which was fully active with NFCD. Using the system contained ME-L31ORIQ401C and DLDH-V152R, we showed that the biotransformation of L-malate into D-Lactate could be done in the presence of a catalytic amount of NFCD, that is to say, NFCD was recyclable by two engineered enzyme. These bioorthogonal systems should be useful in terms of developing bioanalytical methods and producing biochemicals. Our approach opened the window to engineer bioorthogonal redox systems, which will provide us unique tools for systems biology and synthetic biology researches. We will discuss this in more detail during the conference.; Biological redox chemistry is catalyzed by numerous enzymes depending largely on a few cofactors. Nicotinamide adenine dinucleotide (NAD) is one of the most important cofactors, is also involved in diverse non-redox processes. It remains challenging to disconnect one redox reaction from the other NAD-dependent cellular events. 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 multi-site saturated mutagenesis libraries of the NAD-dependent malic enzyme (ME) from Escherichia coli, we identified the mutant ME-L3I0R/Q401C, activity of which was excellent with NFCD, but marginal with NAD. Moreover, wild type ME showed little activity with NFCD. Based on the insights abstracted from this process, we were able to generate a mutant of D-lactate dehydrogenase (DLDH) from Lacrobacillus helvettcus, DLDH-V152R, which was fully active with NFCD. Using the system contained ME-L31ORIQ401C and DLDH-V152R, we showed that the biotransformation of L-malate into D-Lactate could be done in the presence of a catalytic amount of NFCD, that is to say, NFCD was recyclable by two engineered enzyme. These bioorthogonal systems should be useful in terms of developing bioanalytical methods and producing biochemicals. Our approach opened the window to engineer bioorthogonal redox systems, which will provide us unique tools for systems biology and synthetic biology researches. We will discuss this in more detail during the conference.
Language英语
Document Type会议论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/116040
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhao ZB(赵宗保)
Recommended Citation
GB/T 7714
Zhao ZB,Ji DB,Wang L,et al. Assembling of Bioorthogonal Redox Systems Depending on an NAD Analog[C]. 待补充:待补充,2011:61-0.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[赵宗保]'s Articles
[纪德彬]'s Articles
[王磊]'s Articles
Baidu academic
Similar articles in Baidu academic
[赵宗保]'s Articles
[纪德彬]'s Articles
[王磊]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[赵宗保]'s Articles
[纪德彬]'s Articles
[王磊]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.