中国科学院大连化学物理研究所机构知识库
Advanced  
DICP OpenIR  > 中国科学院大连化学物理研究所  > 会议论文
学科主题: 生物工程
题名: Assembling of Bioorthogonal Redox Systems Depending on an NAD Analog
作者: 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)
会议文集: BITS 2nd Symposium on Enzymes and Biocatalysis-2011
会议名称: BITS 2nd Symposium on Enzymes & Biocatalysis-2011
会议日期: 2011-4-25
出版日期: 2011
会议地点: 大连
通讯作者: 赵宗保
出版者: 待补充
出版地: 待补充
合作性质: 分会口头报告
部门归属: 1816
主办者: 中国医药生物技术协会
摘要: 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.
英文摘要: 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.
语种: 英语
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/116040
Appears in Collections:中国科学院大连化学物理研究所_会议论文

Files in This Item:

There are no files associated with this item.


Recommended Citation:
Zhao ZB,Ji DB,Wang L,et al. Assembling of Bioorthogonal Redox Systems Depending on an NAD Analog[C]. 见:BITS 2nd Symposium on Enzymes & Biocatalysis-2011. 大连. 2011-4-25.
Service
 Recommend this item
 Sava as my favorate item
 Show this item's statistics
 Export Endnote File
Google Scholar
 Similar articles in Google Scholar
 [赵宗保]'s Articles
 [纪德彬]'s Articles
 [王磊]'s Articles
CSDL cross search
 Similar articles in CSDL Cross Search
 [赵宗保]‘s Articles
 [纪德彬]‘s Articles
 [王磊]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
  Add to CiteULike  Add to Connotea  Add to Del.icio.us  Add to Digg  Add to Reddit 
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

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

 

 

Valid XHTML 1.0!
Powered by CSpace