Expression of inulinase gene in the oleaginous yeast Yarrowia lipolytica and single cell oil production from inulin-containing materials

被引:80
|
作者
Zhao, Chun-Hai [1 ,2 ,3 ]
Cui, Wei [1 ,2 ]
Liu, Xiao-Yan [1 ,2 ]
Chi, Zhen-Ming [1 ,2 ]
Madzak, Catherine [1 ,2 ]
机构
[1] Ocean Univ China, UNESCO, Chinese Ctr Marine Biotechnol, Qingdao, Peoples R China
[2] Ocean Univ China, Inst Marine Biodivers & Evolut, Qingdao, Peoples R China
[3] Shandong Binzhou Vocat Collge, Binzhou, Peoples R China
基金
中国国家自然科学基金;
关键词
Oleaginous yeast; Yarrowia lipolytica; Exo-inulinase; Single cell oil; Gene expression; PICHIA-GUILLIERMONDII; PROTEIN EXPRESSION; SECRETION; CLONING; BATCH;
D O I
10.1016/j.ymben.2010.09.001
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Yarrowia lipolytica ACA-DC 50109 has been reported to be anoleaginous yeast and significant quantities of lipids were accumulated inside the yeast cells. In this study, the INU1 gene encoding exo-inulinase cloned from Kluyveromyces marxianus CBS 6556 was ligated into the expression plasmid pINA1317 and expressed in the cells of the oleaginous yeast. The activity of the inulinase with 6 x His tag secreted by the transformant Z31 obtained was found to be 41.7 U mL(-1) after cell growth for 78 h. After optimization of the medium and cultivation conditions for single cell oil production, the transformant could accumulate 46.3% (w/w) oil from inulin in its cells and cell dry weight was 11.6 gL(-1) within 78 h at the flask level. During the 2-L fermentation, the transformant could accumulate 48.3% (w/w) oil from inulin in its cells and cell dry weight was 13.3 gL(-1) within 78 h while the transformant could accumulate 50.6% (w/w) oil from extract of Jerusalem artichoke tubers in its cells and cell dry weight was 14.6 gL(-1) with in 78 h. At the end of fermentation, most of the added sugar was utilized by the transformant cells. Over 91.5% of the fatty acids from the transformant cultivated in the extract of Jerusalem artichoke tubercles was C-16:0, C-18:1 and C-18:2, especially C-18:1 (58.5%). (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码:510 / 517
页数:8
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