title: Phosphate-responsive promoter of a Pichia pastoris sodium phosphate symporter.
abstract: 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 (P(PHO89)) were investigated using a bacterial lipase gene as a reporter in 5-liter jar fermentation experiments. P(PHO89) 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 1alpha and the glyceraldehyde-3-phosphate dehydrogenase promoter, P(PHO89) exhibited strong transcriptional activity with higher specific productivity (amount of lipase produced/cell/h). Furthermore, a cost-effective and simple P(PHO89)-based fermentation process was developed for industrial application. These results demonstrate the potential for efficient use of P(PHO89) for controlled production of recombinant proteins in P. pastoris.
authors.affiliation: Biotechnology Process Engineering Center, KRIBB, Daejeon, Republic of Korea.
authors.forename: Jungoh
authors.initials: J
authors.lastname: Ahn
authors.forename: Jiyeon
authors.initials: J
authors.lastname: Hong
authors.forename: Myongsoo
authors.initials: M
authors.lastname: Park
authors.forename: Hyeokweon
authors.initials: H
authors.lastname: Lee
authors.forename: Eungyo
authors.initials: E
authors.lastname: Lee
authors.forename: Chunsuk
authors.initials: C
authors.lastname: Kim
authors.forename: Joohwan
authors.initials: J
authors.lastname: Lee
authors.forename: Eui-sung
authors.initials: ES
authors.lastname: Choi
authors.forename: Joon-ki
authors.initials: JK
authors.lastname: Jung
authors.forename: Hongweon
authors.initials: H
authors.lastname: Lee