Improving the Secretory Expression of an - Galactosidase from Aspergillus niger in Pichia pastoris

doi:10.1371/journal.pone.0161529

QIBEBT-IR > 酶工程研究组

	Improving the Secretory Expression of an - Galactosidase from Aspergillus niger in Pichia pastoris
	Zheng, Xianliang 1,2,3,4; Fang, Bo 1,2,3; Han, Dongfei 1,2; Yang, Wenxia1,2 ; Qi, Feifei1,2 ; Chen, Hui 1,2; Li, Shengying 1,2
	2016-08-22
发表期刊	PLOS ONE
卷号	11 期号:8
摘要	alpha-Galactosidases are broadly used in feed, food, chemical, pulp, and pharmaceutical industries. However, there lacks a satisfactory microbial cell factory that is able to produce alpha-galactosidases efficiently and cost-effectively to date, which prevents these important enzymes from greater application. In this study, the secretory expression of an Aspergillus niger alpha-galactosidase (AGA) in Pichia pastoris was systematically investigated. Through codon optimization, signal peptide replacement, comparative selection of host strain, and saturation mutagenesis of the P1' residue of Kex2 protease cleavage site for efficient signal peptide removal, a mutant P. pastoris KM71H (Mut(s)) strain of AGA-I with the specific P1' site substitution (Glu to Ile) demonstrated remarkable extracellular a-galactosidase activity of 1299 U/ ml upon a 72 h methanol induction in 2.0 L fermenter. The engineered yeast strain AGA-I demonstrated approximately 12-fold higher extracellular activity compared to the initial P. pastoris strain. To the best of our knowledge, this represents the highest yield and productivity of a secreted alpha-galactosidase in P. pastoris, thus holding great potential for industrial application.
文章类型	Article
WOS标题词	Science & Technology
DOI	10.1371/journal.pone.0161529
关键词[WOS]	HIGH-LEVEL EXPRESSION ; HUMAN ALPHA-GALACTOSIDASE ; ALCOHOL OXIDASE ; SACCHAROMYCES-CEREVISIAE ; THERMOMYCES-LANUGINOSUS ; METHYLOTROPHIC YEAST ; CLEAVAGE SITES ; GENE ; CLONING ; PURIFICATION
收录类别	SCI
语种	英语
WOS研究方向	Science & Technology - Other Topics
项目资助者	National Natural Science Foundation of China(NSFC 21406250 ; Applied Basic Research Programs of Science and Technology of Qingdao(15-9-1-106-jch) ; Opening Fund of Key Laboratory of Protein Chemistry and Developmental Biology of the Ministry of Education at Hunan Normal University(2015DF05) ; NSFC 31300663)
WOS类目	Multidisciplinary Sciences
WOS记录号	WOS:000381768400066
引用统计
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/8485
专题	酶工程研究组
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Synthet Biol, 189 Songling Rd, Qingdao 266101, Shandong, Peoples R China 2.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biofuels, 189 Songling Rd, Qingdao 266101, Shandong, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Sinodanish Ctr Educ & Res, Beijing 100190, Peoples R China
推荐引用方式 GB/T 7714	Zheng, Xianliang,Fang, Bo,Han, Dongfei,et al. Improving the Secretory Expression of an - Galactosidase from Aspergillus niger in Pichia pastoris[J]. PLOS ONE,2016,11(8).
APA	Zheng, Xianliang.,Fang, Bo.,Han, Dongfei.,Yang, Wenxia.,Qi, Feifei.,...&Li, Shengying.(2016).Improving the Secretory Expression of an - Galactosidase from Aspergillus niger in Pichia pastoris.PLOS ONE,11(8).
MLA	Zheng, Xianliang,et al."Improving the Secretory Expression of an - Galactosidase from Aspergillus niger in Pichia pastoris".PLOS ONE 11.8(2016).