Effect of Co-expressing Protein HAC1 on Expression of α-galactosidase in Pichia pastoris

Abstract

Abstract: To improve the production of a recombinant α-galactosidase in Pichia pastoris, the hac1 gene from P. pastoris GS115 constructed in pPic9K vector was coexpressed with α-galactosidase gene in P. pastoris X33. Results showed that the HAC1 protein under control of AOX1 promoter increased the expression level of α-galactosidase protein in P. pastoris. The α-galactosidase activity superexpressed with the HAC1 protein was up to 2.2 times than that of constitutive expression of HAC1. hac1 gene integrated into the genome of the P.pastoris was proved by PCR. The galactosidase activity of Gal-HAC1-4#, after 168 h methanol induction in 50 L high-density fermentation, reached the highest value of 6 560 U/mL, and increased by 27% compared with Gal-21#. The crude protein concentration of Gal-HAC1-4# was higher than the strain Gal-21# after methanol induced. These results showed that the galactosidase expression ability of Pichia pastoris was inproved by co-expressing protein HAC1.

Key words: HAC1, α-galactosidase, P. pastoris, Co-exprssion, UPR

Zhong Kaixin, Chen Lizhi, Li Yangyuan. Effect of Co-expressing Protein HAC1 on Expression of α-galactosidase in Pichia pastoris[J]. Biotechnology Bulletin, 2014, 0(7): 184-189.

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