Cell-surface display of the active mannanase in Yarrowia lipolytica with a novel surface-display system.
A novel surface-display system was constructed using the cell-wall anchor protein Flo1p from Saccharomyces cerevisiae, the mannanase (man1) from Bacillus subtilis fused with the C-terminus of Flo1p and the 6xHis tag was inserted between Flo1p and man1. The fusion protein was displayed on the cell surface of Yarrowia lipolytica successfully, and it was confirmed by immunofluorescence. In succession, the surface-displayed mannanase was characterized. The optimum catalytic conditions for the recombinant mannanase were 55 degrees C at pH 6.0, and it exhibited high stability against pH variation. The highest activity of the recombinant mannanase reached 62.3 IU/g (dry cell weight) after the recombinant was cultivated for 96 h in YPD medium [1% (w/v) yeast extract/2% (w/v) peptone/2% (w/v) glucose]. To our knowledge, the present paper is the first to report that high-activity mannanase is displayed on the cell surface of Y. lipolytica with Flo1p.
Yang
Xiao-Song
XS
College of Life Science, Hubei University, Wuhan 430062, People's Republic of China.
Jiang
Zheng-Bing
ZB
Song
Hui-Ting
HT
Jiang
Si-Jing
SJ
Madzak
Catherine
C
Ma
Li-Xin
LX
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D006639:Histidine / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009842:Oligopeptides / Q000235:genetics; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D013696:Temperature; D025062:Yarrowia / Q000235:genetics* / Q000254:growth & development; D044902:beta-Mannosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D003470:Culture Media; C471213:His-His-His-His-His-His; D008565:Membrane Proteins; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D006639:Histidine; D044902:beta-Mannosidase