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Bacterial surface display of a co-factor containing enzyme, ω-transaminase from Vibrio fluvialis using the Bacillus subtilis spore display system.
- Source :
-
Bioscience, biotechnology, and biochemistry [Biosci Biotechnol Biochem] 2011; Vol. 75 (9), pp. 1862-5. Date of Electronic Publication: 2011 Sep 07. - Publication Year :
- 2011
-
Abstract
- To improve the conventional bacterial surface display systems and to display a co-factor containing enzyme, ω-transaminase from Vibrio fluvialis, which needs pyridoxal phosphate (PLP) for efficient transamination, Bacillus subtilis spore display system with cotG, as an anchoring motif was used. Flow cytometry of the B. subtilis spore-expressing ω-transaminase proved its surface localization on the spore. The enzymatic activity of the spore expressing ω-transaminase was more than 30 times higher than that of the host spore. Protease treatment of the ω-transaminase displaying spores resulted in decreased transaminase activity, which is in keeping with the surface location of the fusion protein, CotG-ω-transaminase.
- Subjects :
- Bacillus subtilis enzymology
Bacterial Proteins genetics
Biological Assay
Dimerization
Flow Cytometry
Protein Folding
Pyridoxal Phosphate metabolism
Recombinant Fusion Proteins chemistry
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Spores, Bacterial enzymology
Transaminases chemistry
Transaminases genetics
Transaminases metabolism
Vibrio chemistry
Bacillus subtilis genetics
Bacterial Proteins metabolism
Protein Engineering methods
Recombinant Fusion Proteins isolation & purification
Spores, Bacterial genetics
Transaminases isolation & purification
Vibrio enzymology
Subjects
Details
- Language :
- English
- ISSN :
- 1347-6947
- Volume :
- 75
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Bioscience, biotechnology, and biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 21897008
- Full Text :
- https://doi.org/10.1271/bbb.110307