Expression of inulinase gene in the oleaginous yeast Yarrowia lipolytica and single cell oil production from inulin-containing materials.
Yarrowia lipolytica ACA-DC 50109 has been reported to be an oleaginous 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 × His tag secreted by the transformant Z31 obtained was found to be 41.7U 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 g L(-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 g L(-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 g L(-1) within 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%).
Zhao
Chun-Hai
CH
Unesco Chinese Center of Marine Biotechnology and Institute of Marine Biodiversity and Evolution, Ocean University of China, Yushan Road, No. 5, Qingdao, China.
Cui
Wei
W
Liu
Xiao-Yan
XY
Chi
Zhen-Ming
ZM
Madzak
Catherine
C
D057005:Bioengineering; D002241:Carbohydrates / Q000032:analysis; D003001:Cloning, Molecular; D003470:Culture Media; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D005227:Fatty Acids / Q000032:analysis; D005285:Fermentation; D005822:Genetic Vectors; D006026:Glycoside Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D006368:Helianthus / Q000737:chemistry; D007444:Inulin / Q000378:metabolism*; D007716:Kluyveromyces / Q000201:enzymology / Q000235:genetics; D009584:Nitrogen / Q000378:metabolism; D009821:Oils / Q000737:chemistry / Q000378:metabolism*; D010936:Plant Extracts / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic; D014498:Uracil / Q000378:metabolism; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics*
D002241:Carbohydrates; D003470:Culture Media; D004271:DNA, Fungal; D005227:Fatty Acids; D009821:Oils; D010936:Plant Extracts; D014498:Uracil; D007444:Inulin; D006026:Glycoside Hydrolases; C023086:inulinase; D009584:Nitrogen