Surface display of bacterial tyrosinase on spores of Bacillus subtilis using CotE as an anchor protein.
Tyrosinases, copper-containing monooxygenases, are widely used enzymes for industrial, medical, and environmental applications. We report the first functional surface display of Bacillus megaterium tyrosinase on Bacillus subtilis spores using CotE as an anchor protein. Flow Cytometry was used to verify surface expression of tyrosinase on the purified spores. Moreover, tyrosinase activity of the displayed enzyme on B. subtilis spores was monitored in the presence of L-tyrosine (substrate) and CuSO4(inducer). The stability of the spore-displayed tyrosinase was then evaluated after 15 days maintenance of the spores at room temperature, and no significant decrease in the enzyme activity was observed. In addition, the tyrosinase-expressing spores could be repeatedly used with 62% retained enzymatic activity after six times washing with Tris-HCl buffer. This genetically immobilized tyrosinase on the spores would make a new advance in industrial, medical, and environmental applications.
Faculty of Sciences, Department of Biology, University of Isfahan, Isfahan, Iran.
Afrouzossadat
A
Hosseini-Abari
Interdisciplinary Program for Biochemical Engineering and Biotechnology, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Korea.
Byung-Gee
BG
Kim
Interdisciplinary Program for Biochemical Engineering and Biotechnology, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Korea.
Sang-Hyuk
SH
Lee
Faculty of Sciences, Department of Biology, University of Isfahan, Isfahan, Iran.
Giti
G
Emtiazi
Department of Chemical Engineering, College of Engineering, Dong-A University, Busan, Korea.
Wooil
W
Kim
Department of Chemical Engineering, College of Engineering, Dong-A University, Busan, Korea.
June-Hyung
JH
Kim
D001410:Bacillus megaterium / Q000201:enzymology* / Q000235:genetics; D001412:Bacillus subtilis / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D061505:Cell Surface Display Techniques; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000235:genetics* / Q000378:metabolism; D005434:Flow Cytometry; D007218:Industrial Microbiology / Q000379:methods; D014442:Monophenol Monooxygenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013171:Spores, Bacterial / Q000201:enzymology / Q000235:genetics*; D013499:Surface Properties; D014443:Tyrosine / Q000378:metabolism
D001426:Bacterial Proteins; C077494:CotE protein, Bacillus subtilis; D004800:Enzymes, Immobilized; D011993:Recombinant Fusion Proteins; D014443:Tyrosine; D014442:Monophenol Monooxygenase
Bacillus subtilis; CotE; Spore surface display; Tyrosinase