Hwang Bum-Yeol, Kim Byung-Gee, Kim June-Hyung
Department of Chemical Engineering and Bioengineering and The Helen Wills Neuroscience Institute, University of California, Berkeley, CA 94720-3220, USA.
Biosci Biotechnol Biochem. 2011;75(9):1862-5. doi: 10.1271/bbb.110307. Epub 2011 Sep 7.
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.
为改进传统的细菌表面展示系统，并展示一种含有辅因子的酶——来自河流弧菌的ω-转氨酶（其高效转氨作用需要磷酸吡哆醛（PLP）），使用了以cotG作为锚定基序的枯草芽孢杆菌孢子展示系统。对表达ω-转氨酶的枯草芽孢杆菌孢子进行流式细胞术分析，证明其在孢子表面定位。表达ω-转氨酶的孢子的酶活性比宿主孢子高30倍以上。对展示ω-转氨酶的孢子进行蛋白酶处理，导致转氨酶活性降低，这与融合蛋白CotG-ω-转氨酶的表面定位一致。
