Pichia pastoris wall protein gcw51 and its surface display system and construction method

Abstract

The invention discloses Pichia pastoris wall protein, a surface display system constructed thereby and a construction method thereof. The amino acid sequence of the Pichia pastoris wall protein is SEQ NO:1. The Pichia pastoris wall protein cell surface display system is formed by using the Pichia pastoris wall protein Gcw51 as ankyrin and fixing target protein on the surface of the Pichia pastoris cell. The expression quantity of the wall protein Gcw51 in the Pichia pastoris is very high and is respectively 9 times and 21 times higher than those of the endogenesis ankyrin Pir1 protein and Pir2 protein applied to the Pichia pastoris surface display system.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Pichia pastoris wall protein Gcw51 and its constructed surface display system and construction method. Background technique [0002] At present, the host bacteria that are widely used in the microbial surface display system mainly include phages, bacteria (such as Escherichia coli Escherichia. coli Proteus mirabilis Proteus mirabilis etc.), Saccharomyces cerevisiae ( Saccharomyces cerevisiae ) and Pichia pastoris ( Pichia pastoris ). The anchoring proteins used for surface display generally have the following characteristics: (1) they are firmly anchored on the cell surface and will not easily fall off the cell surface; (2) they can be effectively fused with the target protein sequence without affecting the structure of the target protein and function; (3) have certain resistance to protease. The anchor proteins of the bacterial surface display system mainly include bacter...

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Problems solved by technology

However, the inventors found that the expression level of the Pir protein itself is low, and the effect of the anchor protein as the surface display system of Pichia pastoris is general

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