Properties of alkaline protease genetically engineered on cell surface of the yeast Yarrowia lipolytica.
ALP2 gene encoding alkaline protcase cloned from Aureobasidium pullulans HN2-3 was ligated into the surface display plasmid and expressed in the cells of the yeast Yarrowia lipolytica. The expressed alkaline protease was immobilized on the yeast cells. The activity of the immobilized enzyme with 6 x His tag was found to be significantly higher than that of without 6 x His tag. The immobilized enzyme showed lower optimal temperature and a lower affinity for azocasein than the free enzyme purified from A. pullulans HN2-3. The thermal stability of the immobilized enzyme enhanced and the pH stability decreased, compared to that of the free enzyme.
Unesco Chinese Center of Marine Biotechnology, Ocean University of China, Yushan Road, No. 5, Qingdao, China.
Xiumei
X
Ni
Lixi
L
Yue
Jing
J
Li
Zhenming
Z
Chi
Zhiqiang
Z
Liu
Catherine
C
Madzak
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D002364:Caseins / Q000737:chemistry; D002412:Cations; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics*; D004800:Enzymes, Immobilized / Q000737:chemistry; D005658:Fungi / Q000201:enzymology; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007700:Kinetics; D008954:Models, Biological; D013696:Temperature; D025062:Yarrowia / Q000201:enzymology / Q000235:genetics*
D001426:Bacterial Proteins; D002364:Caseins; D002412:Cations; D004800:Enzymes, Immobilized; D007477:Ions; C026018:azocasein; D010450:Endopeptidases; C087822:alkaline protease