Preparation of a whole-cell biocatalyst of Aspergillus niger lipase and its practical properties.
Aspergillus niger lipase (ANL), a widely used hydrolase, was displayed for the first time on the surface of Saccharomyces cerevisiae using a-agglutinin as an anchor protein. Localization of ANL on the cell surface was confirmed by immunofluorescence microscopy. The displayed ANL was confirmed to be active toward tributyrin and p-nitrophenyl caprylate (pNPC). The hydrolytic activity toward pNPC reached 43.8 U/g of dry cell weight after induction by galactose for 72 h. The ANL-displaying cells were characterized for their use as whole-cell biocatalysts. The optimum temperature was 45 °C, and the pH was 7.0. The cells had good thermostability, retaining almost 80% of the full activity after incubation at 60 °C for 1 h, and >80% of the full activity at 50 °C for 6 h. The displayed lipase showed a preference for medium-chain fatty acid p-nitrophenyl esters. Therefore, the produced whole-cell catalyst is likely to have a wide range of applications.
Liu
Wenshan
W
Key Laboratory of Molecular Biophysics, Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
Jia
Bin
B
Zhao
Heyun
H
Xu
Li
L
Yan
Yunjun
Y
D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D002384:Catalysis; D002473:Cell Wall / Q000201:enzymology; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D014162:Transfection
D011994:Recombinant Proteins; D008049:Lipase