Production of l-alanyl-l-glutamine by immobilized Pichia pastoris GS115 expressing -amino acid ester acyltransferase

doi:10.1186/s12934-019-1077-1

	Production of l-alanyl-l-glutamine by immobilized Pichia pastoris GS115 expressing -amino acid ester acyltransferase
	Gao, Jiao-Qi 2; Li, Yi-Min 1; Pei, Xu-Ze 1; Du, Cong 1; Fan, Chao 3; Yuan, Wen-Jie 1; Bai, Feng-Wu 4
刊名	MICROBIAL CELL FACTORIES
	2019-02-02
卷号	18 页码:12
关键词	l-Alanyl-l-glutamine -Amino acid ester acyltransferase Immobilization Codon optimization Recycle
ISSN号	1475-2859
DOI	10.1186/s12934-019-1077-1
通讯作者	Yuan, Wen-Jie(ywj@dlut.edu.cn)
英文摘要	Backgroundl-Alanyl-l-glutamine (Ala-Gln) represents the great application potential in clinic due to the unique physicochemical properties. A new approach was developed to synthesize Ala-Gln by recombinant Escherichia coli OPA, which could overcome the disadvantages of traditional chemical synthesis. Although satisfactory results had been obtained with recombinant E. coli OPA, endotoxin and the use of multiple antibiotics along with toxic inducer brought the potential biosafety hazard for the clinical application of Ala-Gln.ResultsIn this study, the safer host Pichia pastoris was applied as an alternative to E. coli. A recombinant P. pastoris (named GPA) with the original gene of -amino acid ester acyltransferase (SsAet) from Sphingobacterium siyangensis SY1, was constructed to produce Ala-Gln. To improve the expression efficiency of SsAet in P. pastoris, codon optimization was conducted to obtain the strain GPAp. Here, we report that Ala-Gln production by GPAp was approximately 2.5-fold more than that of GPA. The optimal induction conditions (cultivated for 3days at 26 degrees C with a daily 1.5% of methanol supplement), the optimum reaction conditions (28 degrees C and pH 8.5), and the suitable substrate conditions (AlaOMe/Gln=1.5/1) were also achieved for GPAp. Although most of the metal ions had no effects, the catalytic activity of GPAp showed a slight decrease in the presence of Fe3+ and an obvious increase when cysteine or PMSF were added. Under the optimum conditions, the Ala-Gln generation by GPAp realized the maximum molar yield of 63.5% and the catalytic activity of GPAp by agar embedding maintained extremely stable after 10 cycles.ConclusionsCharacterized by economy, efficiency and practicability, production of Ala-Gln by recycling immobilized GPAp (whole-cell biocatalyst) is represents a green and promising way in industrial.
资助项目	projects of International Cooperation and Exchanges NSFC[51561145014] ; Dalian Innobio Corporation Limited[HX20160536]
WOS关键词	DIPEPTIDES ; STABILITY ; GLUCOSE
WOS研究方向	Biotechnology & Applied Microbiology
语种	英语
出版者	BMC
WOS记录号	WOS:000457553500001
资助机构	projects of International Cooperation and Exchanges NSFC ; projects of International Cooperation and Exchanges NSFC ; Dalian Innobio Corporation Limited ; Dalian Innobio Corporation Limited ; projects of International Cooperation and Exchanges NSFC ; projects of International Cooperation and Exchanges NSFC ; Dalian Innobio Corporation Limited ; Dalian Innobio Corporation Limited ; projects of International Cooperation and Exchanges NSFC ; projects of International Cooperation and Exchanges NSFC ; Dalian Innobio Corporation Limited ; Dalian Innobio Corporation Limited ; projects of International Cooperation and Exchanges NSFC ; projects of International Cooperation and Exchanges NSFC ; Dalian Innobio Corporation Limited ; Dalian Innobio Corporation Limited
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/166112]
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	Yuan, Wen-Jie
作者单位	1.Dalian Univ Technol, Sch Life Sci & Biotechnol, Dalian 116024, Peoples R China 2.Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China 3.Dalian Innobio Corp Ltd, Res & Dev Ctr, Dalian 116600, Peoples R China 4.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China
推荐引用方式 GB/T 7714	Gao, Jiao-Qi,Li, Yi-Min,Pei, Xu-Ze,et al. Production of l-alanyl-l-glutamine by immobilized Pichia pastoris GS115 expressing -amino acid ester acyltransferase[J]. MICROBIAL CELL FACTORIES,2019,18:12.
APA	Gao, Jiao-Qi.,Li, Yi-Min.,Pei, Xu-Ze.,Du, Cong.,Fan, Chao.,...&Bai, Feng-Wu.(2019).Production of l-alanyl-l-glutamine by immobilized Pichia pastoris GS115 expressing -amino acid ester acyltransferase.MICROBIAL CELL FACTORIES,18,12.
MLA	Gao, Jiao-Qi,et al."Production of l-alanyl-l-glutamine by immobilized Pichia pastoris GS115 expressing -amino acid ester acyltransferase".MICROBIAL CELL FACTORIES 18(2019):12.