Carbon and nitrogen sources modulate lipase production in the yeast Yarrowia lipolytica

被引:98
作者
Fickers, P
Nicaud, JM
Gaillardin, C
Destain, J
Thonart, P
机构
[1] Univ Liege, Serv Technol Microbienne, Ctr Wallon Biol Ind, B-4000 Liege, Belgium
[2] Inst Natl Agron Paris Grignon, CNRS, UMR2585, Lab Microbiol & Genet Mol, Thiverval Grignon, France
[3] Fac Univ Sci Agron Gembloux, Ctr Wallon Biol Ind, Unite Bioindustries, Gembloux, Belgium
关键词
enzyme location; histidine tag; LIP2; regulation; lipase; Yarrowia lipolytica;
D O I
10.1111/j.1365-2672.2004.02190.x
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
Aims: To analyse the influence of nitrogen and carbon sources on extracellular lipase production by Yarrowia lipolytica-overproducing mutant in order to optimize its production in large-scale bioreactors. Methods and Results: The level of lipase production and LIP2 induction, measured using an LIP2-LacZ reporter gene, were compared for different carbon and nitrogen sources and for different concentrations. The localization of the enzyme during growth was also determined by Western blotting analysis using a six-histidine-tagged lipase. Significance and Impact of the Study: Tryptone N1 and oleic acid are the most suitable nitrogen and carbon sources for the production of the extracellular lipase by the Y. lipolytica mutant. Higher levels of lipase production were obtained as the tryptone concentration increased in the culture medium. Such a positive correlation was not observed with oleic acid media where the highest lipolytic productivities were obtained in the presence of low concentration. We also demonstrate that in the presence of oleic acid, lipase is cell-bound during the growth phase before being released in the media. Conclusions: This work provides a better understanding of the mechanism controlling LIP2 expression and, thus, extracellular lipase production in the yeast Y. lipolytica.
引用
收藏
页码:742 / 749
页数:8
相关论文
共 23 条
[1]  
Barth G, 1996, NONCONVENTIONAL YEAS, P313
[2]   Identification of a triacylglycerol lipase gene family in Candida deformans:: molecular cloning and functional expression [J].
Bigey, F ;
Tuery, K ;
Bougard, D ;
Nicaud, JM ;
Moulin, G .
YEAST, 2003, 20 (03) :233-248
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   EXTRACELLULAR RNASE PRODUCED BY YARROWIA-LIPOLYTICA [J].
CHENG, SC ;
OGRYDZIAK, DM .
JOURNAL OF BACTERIOLOGY, 1986, 168 (02) :581-589
[5]   Production and characteristics of the lipase from Yarrowia lipolytica 681 [J].
Corzo, G ;
Revah, S .
BIORESOURCE TECHNOLOGY, 1999, 70 (02) :173-180
[6]   Improvement of lipase production from Yarrowia lipolytica [J].
Destain, J ;
Roblain, D ;
Thonart, P .
BIOTECHNOLOGY LETTERS, 1997, 19 (02) :105-107
[7]   Overproduction of lipase by Yarrowia lipolytica mutants [J].
Fickers, P ;
Nicaud, JM ;
Destain, J ;
Thonart, P .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2003, 63 (02) :136-142
[8]   LEU2 DIRECTED EXPRESSION OF BETA-GALACTOSIDASE ACTIVITY AND PHLEOMYCIN RESISTANCE IN YARROWIA-LIPOLYTICA [J].
GAILLARDIN, C ;
RIBET, AM .
CURRENT GENETICS, 1987, 11 (05) :369-375
[9]   BACTERIAL LIPASES [J].
JAEGER, KE ;
RANSAC, S ;
DIJKSTRA, BW ;
COLSON, C ;
VANHEUVEL, M ;
MISSET, O .
FEMS MICROBIOLOGY REVIEWS, 1994, 15 (01) :29-63
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+