The Pichia pastoris transmembrane protein GT1 is a glycerol transporter and relieves the repression of glycerol on AOX1 expression

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Zhan, Chunjun and Wang, Songwei and Sun, Yang and Dai, Xiaofeng and Liu, Xiuxia and Harvey, Linda and McNeil, Brian and Yang, Yankun and Bai, Zhonghu (2016) The Pichia pastoris transmembrane protein GT1 is a glycerol transporter and relieves the repression of glycerol on AOX1 expression. FEMS Yeast Research, 16 (4). pp. 1-10. ISSN 1567-1364 (https://doi.org/10.1093/femsyr/fow033)

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Abstract

Promoter of alcohol oxidase I (PAOX1) is the most efficient promoter involved in the regulation of recombinant protein expression in Pichia pastoris (P. pastoris). PAOX1 is tightly repressed by the presence of glycerol in the culture medium; thus, glycerol must be exhausted before methanol can be taken up by P. pastoris and the expression of the heterologous protein can be induced. In this study, a candidate glycerol transporter (GT1, GeneID: 8197545) was identified, and its role was confirmed by further studies (e.g. bioinformatics analysis, heterologous complementation in Schizosaccharomyces pombe (S. pombe)). When GT1 is co-expressed with enhanced green fluorescent protein (EGFP), it localizes to the membrane and S. pombe carrying gt1 but not the wild-type strain can grow on medium containing glycerol as the sole carbon source. The present study is the first to report that AOX1 in the X-33gt1 mutant can achieve constitutive expression in medium containing glycerol; thus, knocking down gt1 can eliminate the glycerol repression of PAOX1 in P. pastoris. These results suggest that the glycerol transporter may participate in the process of PAOX1 inhibition in glycerol medium.

  • Item type:Article
    ID code:60123
    Dates:
    Date
    Event
    1 June 2016
    Published
    11 April 2016
    Published Online
    1 April 2016
    Accepted
    Notes:This is a pre-copyedited, author-produced version of an article accepted for publication in FEMS Yeast Research following peer review. Zhan, C., Wang, S., Sun, Y., Dai, X., Liu, X., Harvey, L., ... Bai, Z. (2016). The Pichia pastoris transmembrane protein GT1 is a glycerol transporter and relieves the repression of glycerol on AOX1 expression. FEMS Yeast Research, 16(4), 1-10. DOI: 10.1093/femsyr/fow033
    Subjects:Medicine > Pharmacy and materia medica
    Department:Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
    Depositing user: Pure Administrator
    Date deposited:09 Mar 2017 11:59
    Last modified:11 Nov 2024 11:27
    Related URLs:
    URI:https://strathprints.strath.ac.uk/id/eprint/60123
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