High-level expression of a ZEN-detoxifying gene by codon optimization and biobrick in Pichia pastoris

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	High-level expression of a ZEN-detoxifying gene by codon optimization and biobrick in Pichia pastoris
	Xiang, La 1; Wang, Qinhong 2; Zhou, Yuling 1; Yin, Lifeng 1; Zhang, Guimin 1; Ma, Yanhe 2
刊名	MICROBIOLOGICAL RESEARCH
	2016
卷号	193 页码:48-56
关键词	Zearalenone degradation Biobrick Protein expression High-density fermentation
英文摘要	The mycotoxin zearalenone (ZEN) can be degraded by a lactone hydrolase ZHD, which was derived from Gliocladium roseum. Here, based on the native ZHD encoding gene zhd101, a codon optimized zhd gene was synthesized, which was used for high expression of ZHD in Pichia pastoris GS115. Meanwhile, to further improve the expression of recombinant ZHD, the plasmids containing 1 to 4 copies of the zhd expression cassette were constructed, respectively, using the biobrick method. The protein expression in the recombinant P. pastoris X3c, which was transformed with the plasmid containing 3 copies of zhd expression cassette, was the highest. In addition, the enzymatic activity of ZHD against ZEN was defined for the first time based on a standard curve of peak area vs ZEN concentration. The ZEN degradation activity of ZHD from shake flask fermentation was calculated as 22.5 U/mL with the specific activity of 4976.5 U/mg. Furthermore, the high-density fermentation of P. pastoris X3c strain was also performed in 5 L fermenter. The maximum enzyme activity of the supernatant was 150.1 U/mL, which were 6.7-fold higher than that of the shake flask fermentation. (C) 2016 Published by Elsevier GmbH.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Microbiology
研究领域[WOS]	Microbiology
关键词[WOS]	ZEARALENONE ; DETOXIFICATION ; MYCOTOXIN ; CORN ; METABOLISM ; STABILITY ; COMPOUND ; FOOD
收录类别	SCI
语种	英语
WOS记录号	WOS:000388247400006
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/2293]
专题	天津工业生物技术研究所_进化代谢工程和分子进化研究组王钦宏_期刊论文
作者单位	1.Hubei Univ, Coll Life Sci, Hubei Collaborat Innovat Ctr Green Transformat Bi, Wuhan 430062, Peoples R China 2.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China
推荐引用方式 GB/T 7714	Xiang, La,Wang, Qinhong,Zhou, Yuling,et al. High-level expression of a ZEN-detoxifying gene by codon optimization and biobrick in Pichia pastoris[J]. MICROBIOLOGICAL RESEARCH,2016,193:48-56.
APA	Xiang, La,Wang, Qinhong,Zhou, Yuling,Yin, Lifeng,Zhang, Guimin,&Ma, Yanhe.(2016).High-level expression of a ZEN-detoxifying gene by codon optimization and biobrick in Pichia pastoris.MICROBIOLOGICAL RESEARCH,193,48-56.
MLA	Xiang, La,et al."High-level expression of a ZEN-detoxifying gene by codon optimization and biobrick in Pichia pastoris".MICROBIOLOGICAL RESEARCH 193(2016):48-56.