[Cell surface display of Yarrowia lipolytica lipase Lip2 in Saccharomyces cerevisiae with a-agglutinin as carrier protein].
OBJECTIVE
In order to display extracellular.lipase Lip2 from Yarrowia lipolytica on the surface of yeast Saccharomyces cerevisiae for whole cell catalysts.


METHODS
The mature Lip2 encoding fragment was amplified from Yarrowia lipolytica total DNA, and was inserted into the 3'terminal of AGA2 to give the plasmid pCTLIP2 for surface display of Lip2. Olive oil, tributyrin and p-nitrophenyl palmitate (pNPP) were used as substrates to measure lipase activity. Moreover, the characterization of displayed lipase and its free form was analyzed.


RESULTS
The surface displayed lipase was confirmed to be active towards olive oil, tributyrin and p-nitrophenyl palmitate (pNPP), and reached its highest expression level at 182 U/g dry cell after induced by galactose for 72h. The optimum temperature of cell surface displayed Lip2 was 40 degrees C After incubated at 50 degrees C for 4h, the surface displayed lipase retained 23.2% of its full activity, improved a little compared to free Lip2. The surface displayed lipase showed a preference to medium-chain and long-chain fatty acids p-nitrophenyl esters (C8-C16).


CONCLUSIONS
The cell surface display system based on a-agglutinin is an effective system for displaying Lip2, and the whole cell EBY100-pCTLIP2 will be probably suited to a different range of applications.
Liu
Wenshan
W
College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China. lwenshan@yahoo.com.cn
Xu
Li
L
Zhao
Heyun
H
Yang
Jiangke
J
Yan
Yunjun
Y
D000373:Agglutinins / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry; D003470:Culture Media; D004800:Enzymes, Immobilized / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D007218:Industrial Microbiology; D008049:Lipase / Q000235:genetics* / Q000378:metabolism; D000069463:Olive Oil; D010938:Plant Oils / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014280:Triglycerides / Q000737:chemistry; D025062:Yarrowia / Q000201:enzymology / Q000235:genetics*
D000373:Agglutinins; D003470:Culture Media; D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D000069463:Olive Oil; D010938:Plant Oils; D011993:Recombinant Fusion Proteins; D014280:Triglycerides; D008049:Lipase; C005830:tributyrin