Bacterial Surface Display of Levansucrase of Zymomonas mobilis Using Bacillus Subtilis Spore Display System
2011
Kim, J.H., Dong-A University, Busan, Republic of Korea | Choi, S.K., Systems Microbiology Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea | Jung, H.C., Systems Microbiology Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea | Pan, J.G., Systems Microbiology Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea | Kim, B.G., Seoul National University, Seoul, Republic of Korea
Using Bacillus subtilis spore display system, with cotG as an anchoring motif, levansucrase from Zymomonas mobilis, was displayed on the outer surface of Bacillus subtilis spore. Flow cytometry of DB104 (pSDJH-cotG-levU) spore, proved the surface localization of CotG-LevU fusion protein on the spore compared to that of DB104. Enzymatic activity of DB104 (pSDJH-cotG-levU) spore showed more than 1.5 times higher levansucrase specific activity compared to that of the host spore, which is a remarkable increase of enzymatic activity considering the existence of sacA (sucrase) and sacB (levansucrase) in the Bacillus subtilis chromosome. The spore integrity, revealed by sporulation frequency test after heat and lysozyme treatment of spore, did not changed at all in spite of the CotG-LevU fusion protein incorporation into the spore coat layer during spore formation process. These data prove again that Bacillus subtilis spore could be considered as good live immobilization vehicle for efficient bioconversion process.
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