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| The Disruption of STE13 Gene in Pichia pastoris Improves the Expression and Bioactivity of GGH |
| LU Qing-peng, XU Zhen-hong, SI Jin-song, DOU Wen-fang |
| Laboratory of Pharmaceutical Engineering, School of Pharmaceutical Sciences, Jiangnan University, Wuxi 214122, China |
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Abstract Objective: It is barely found that the bioactivity of recombination protein (GLP-1 A2G ) 2 -HSA (GGH) by the cell bioactivity evaluation that related to its protein degradation, which expressed in methylotrophic yeast Pichia pastoris GS115 strain. Therefore, A new P.pastoris GS115 strain with STE13 gene disruption was engineered, helped for the expression of intact GGH, and the bioactivity can be further improved. Methods: The STE13 gene deficient strain GS115/D13 was constructed by the disruption of the P.pastoris homolog of the Saccharomyces cereviaiae dipeptidyl aminopeptidase (STE13 ) gene in Pichia genome by the Cre-Loxp recombination system based on the fusion PCR technology, and the bioactivity of GGH estimated by the cell biological evaluation methods. Results: The recombinant strain GS115/D13 showed the protein degradation much more relieved, apparent bioactivity improved and the yield of about 25.8% increased compared with that in the wild-type strain (GS115/W). Conclusion: The results suggested that STE13 gene disruption offered an expression host for the product of much more intact GGH and helped improve the protein bioactivity.
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Received: 14 January 2013
Published: 25 April 2013
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