Phosphate-Responsive Promoter of a Pichia pastoris Sodium Phosphate Symporter
2009
Ahn, Jungoh | Hong, Jiyeon | Park, Myongsoo | Lee, Hyeokweon | Lee, Eungyo | Kim, Chunsuk | Lee, Joohwan | Choi, Eui-sung | Jung, Joon-ki | Lee, Hongweon
To develop a functional phosphate-regulated promoter in Pichia pastoris, a phosphate-responsive gene, PHO89, which encodes a putative sodium (Na⁺)-coupled phosphate symporter, was isolated. Sequencing analyses revealed a 1,731-bp open reading frame encoding a 576-amino-acid polypeptide with 12 putative transmembrane domains. The properties of the PHO89 promoter (PPHO₈₉) were investigated using a bacterial lipase gene as a reporter in 5-liter jar fermentation experiments. PPHO₈₉ was tightly regulated by phosphate and was highly activated when the cells were grown in a phosphate-limited external environment. Compared to translation elongation factor 1α and the glyceraldehyde-3-phosphate dehydrogenase promoter, PPHO₈₉ exhibited strong transcriptional activity with higher specific productivity (amount of lipase produced/cell/h). Furthermore, a cost-effective and simple PPHO₈₉-based fermentation process was developed for industrial application. These results demonstrate the potential for efficient use of PPHO₈₉ for controlled production of recombinant proteins in P. pastoris.
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