Display of heterologous protein on the surface of Lactobacillus plantarum by using the CspI anchor protein

作者：Xu Wensheng; Zhang Yanyan; Wang Xiaodong; Huang Manqing; Gao Xiuzhi; Zhang Dage; Li Chan; Ai Qijun^*

来源：Annals of Microbiology, 2013, 63(3): 855-861.

DOI：10.1007/s13213-012-0536-2

• Lactobacillus plantarum
• Cell surface protein
• LPxTG motif
• CspI

The C-terminus of the putative cell surface protein CspI which contains one putative LPxTG motif region and a signal peptides fragment were amplified from L. plantarum CICC6024, and the green fluorescent protein gene gfp was amplified from the plasmid pACGFP. The three genes were ligated and the fusion gene was named SgfpL. The fusion gene SgfpL was then cloned into shuttle expression vector pMG36e and transformed into L. plantarum. SDS-PAGE identified that the fusion protein was expressed and the band of fusion protein was observed at the predicated molecular size. Fluorescence assay, western blot against GFP antibody, protease accessibility and SDS sensitivity assays were performed to determine that the GFP was successfully displayed on the surfaces of L. plantarum cells and the maximum display capacity of the GFP fusion protein was ca. 65 mu g ml(-1). The fermentation condition experiments determined that the amounts of GFP fusion protein were increased at a higher temperature and reached the peak at 2.5 h. Then, the beta-galactosidase from Bifidobacterium bifidum was functionally displayed on the surface of L. plantarum cells via CspI to demonstrate the applicability of the CspI-mediated surface display system.

• 出版日期2013-9
• 单位中国农业大学