Effect of Glycosylation of Brucella P39 Antigen in Pichia pastoris on Macrophage Phagocytosis

doi:10.13560/j.cnki.biotech.bull.1985.2018-0170

Abstract

Abstract: P39 protein is one of the most predominant antigenic proteins identified from Brucella by proteomics. First,the dominant antigen protein gene P39 was expressed in Pichia pastoris,and the expressed protein was purified and identified by Western blot. Then,the glycosylation of the protein was identified by NMR（nuclear magnetic resonance）conjugated PNGaseF glycosylase,and the interaction speed between the protein and macrophage cell was studied. The results showed that P39 protein was secreted in P. pastoris GS115 and the expression level was up to 5.05 g/L. NMR results demonstrated that the recombinant P39 protein was modified by glycosylation. The macrophage capture effect of the modified protein was significantly stronger than prokaryotic protein,indicating that the glycosylation modification may affect the efficiency of the interaction between the target protein and the cell,which provides a reference for further exploring the effect of glycosylation on P39 protein and candidate antigen of Brucella glycosylated vaccine.

Key words: Brucella, P39 protein, Pichia pastoris, glycosylation

LIU Si-yang, WANG Yan, FU Yu-qin, HU Xiang-kun, WANG Ming-yu, LU Tian-cheng, WANG Xiu-ran. Effect of Glycosylation of Brucella P39 Antigen in Pichia pastoris on Macrophage Phagocytosis[J]. Biotechnology Bulletin, 2018, 34(9): 237-243.

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