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

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	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, Wenxia 1,2; Qi, Feifei 1,2; Chen, Hui 1,2; Li, Shengying 1,2
刊名	PLOS ONE
	2016-08-22
卷号	11 期号:8
英文摘要	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.
WOS标题词	Science & Technology
类目[WOS]	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
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记录号	WOS:000381768400066
内容类型	期刊论文
源URL	[http://ir.qibebt.ac.cn/handle/337004/8485]
专题	青岛生物能源与过程研究所_酶工程团队
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).