Construction of Pichia pastoris Surface Display Technology as Whole -cell Biocatalyst for Thymidine Phosphorylase

doi:10.13560/j.cnki.biotech.bull.1985.2016.01.032

Abstract

Abstract: Thymidine phosphorylase(TP)extensively involves in nucleoside metabolism and catalyzes the formation of many nucleoside analogs(NA). A yeast cell surface display system for TP was firstly constructed in this study as whole-cell biocatalyst. A deoA gene encoding TP was cloned from Escherichia coli K12 strain and inserted into the yeast expression vector pKFS. Recombinant vector was linearized and electro-transformed into Pichia pastoris GS115 cells. High copy transformant was induced by methanol for 96 h, and the results of immunofluorescence indicated that TP successfully displayed on the surface of P. pastoris. β-thymidine was used as substrate and recombinant GS115 cells as whole-cell biocatalyst, HPLC analysis demonstrated that TP on the surface of P. pastoris had catalytic activity, and catalyzed the production from β-thymidine to thymine.

Key words: thymidine phosphorylase, whole-cell catalysis, Pichia pastoris, surface display

WANG Jie, YU Lei, YANG Dong, LI Jie, WANG Hong-zhong. Construction of Pichia pastoris Surface Display Technology as Whole -cell Biocatalyst for Thymidine Phosphorylase[J]. Biotechnology Bulletin, 2016, 32(1): 201-206.

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