Structural Insights into Phosphite Dehydrogenase Variants Favoring a Non-natural Redox Cofactor

doi:10.1021/acscatal.8b04822

	Structural Insights into Phosphite Dehydrogenase Variants Favoring a Non-natural Redox Cofactor
	Xue, Song 1; Liu, Yuxue 1,2; Feng, Yanbin 1; Wang, Lei 1; Guo, Xiaojia 1,2; Liu, Wujun 1; Li, Qing 1,2; Wang, Xueying 1; Zhao, Zongbao Kent 1,3
刊名	ACS CATALYSIS
	2019-03-01
卷号	9 期号:3 页码:1883-1887
关键词	phosphite dehydrogenase non-natural redox cofactor NAD-binding pocket nicotinamide cytosine dinucleotide cofactor preference
ISSN号	2155-5435
DOI	10.1021/acscatal.8b04822
通讯作者	Xue, Song(xuesong@dicp.ac.cn) ; Zhao, Zongbao Kent(zhaozb@dicp.ac.cn)
英文摘要	Implementation of a non-natural cofactor alternative to the ubiquitous redox cofactor nicotinamide adenosine dinucleotide (NAD) is of great scientific and biotechnological interest. Several redox enzymes have been engineered to favor nicotinamide cytosine dinucleotide (NCD), a smaller-sized NAD analogue. However, molecular interactions involving NAD analogues remain elusive, preventing us from devising more enzymes to accept those analogues. Here we took a semirational approach to evolve phosphite dehydrogenase (Pdh) and identified variants with substantially improved NCD preference. These mutants are valuable components for regeneration of reduced NCD by using phosphite as the electron donor. We then collected X-ray crystal structures of three Pdh variants and their NCD-complexes to delineate molecular basis for NCD binding. It was found that the incorporation of amino acid residues with large side chains enclosing the NAD-binding pocket led to compacted environment favoring NCD over NAD, and additional interactions between NCD and these side chains. These results guided successful engineering of more Pdh mutants with good NCD preference. As many redox enzymes share key structural features, our strategy may be readily adopted to devise NCD-favoring enzymes. We expected that, in the near future, more synthetic systems linked to non-natural cofactors will be created as alternative tools for widespread applications to address challenging problems by chemical and synthetic biologists.
资助项目	National Natural Science Foundation of China[21572227] ; National Natural Science Foundation of China[21778053] ; National Natural Science Foundation of China[21721004] ; Dalian Institute of Chemical Physics, CAS[DICPQIBEBT UN201706] ; Dalian Institute of Chemical Physics, CAS[DICP ZZBS201605]
WOS关键词	EMISSIVE SYNTHETIC COFACTORS ; REGENERATION ; EVOLUTION ; NAD(+) ; ANALOGS ; NADH
WOS研究方向	Chemistry
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000460600600025
资助机构	National Natural Science Foundation of China ; National Natural Science Foundation of China ; Dalian Institute of Chemical Physics, CAS ; Dalian Institute of Chemical Physics, CAS ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Dalian Institute of Chemical Physics, CAS ; Dalian Institute of Chemical Physics, CAS ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Dalian Institute of Chemical Physics, CAS ; Dalian Institute of Chemical Physics, CAS ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Dalian Institute of Chemical Physics, CAS ; Dalian Institute of Chemical Physics, CAS
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/165860]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Xue, Song; Zhao, Zongbao Kent
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Chinese Acad Sci, Dalian Inst Chem Phys, State Key Lab Catalysis, Dalian 116023, Peoples R China
推荐引用方式 GB/T 7714	Xue, Song,Liu, Yuxue,Feng, Yanbin,et al. Structural Insights into Phosphite Dehydrogenase Variants Favoring a Non-natural Redox Cofactor[J]. ACS CATALYSIS,2019,9(3):1883-1887.
APA	Xue, Song.,Liu, Yuxue.,Feng, Yanbin.,Wang, Lei.,Guo, Xiaojia.,...&Zhao, Zongbao Kent.(2019).Structural Insights into Phosphite Dehydrogenase Variants Favoring a Non-natural Redox Cofactor.ACS CATALYSIS,9(3),1883-1887.
MLA	Xue, Song,et al."Structural Insights into Phosphite Dehydrogenase Variants Favoring a Non-natural Redox Cofactor".ACS CATALYSIS 9.3(2019):1883-1887.