Phosphate-responsive promoter of a Pichia pastoris sodium phosphate symporter.
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.
Ahn
Jungoh
J
Biotechnology Process Engineering Center, KRIBB, Daejeon, Republic of Korea.
Hong
Jiyeon
J
Park
Myongsoo
M
Lee
Hyeokweon
H
Lee
Eungyo
E
Kim
Chunsuk
C
Lee
Joohwan
J
Choi
Eui-sung
ES
Jung
Joon-ki
JK
Lee
Hongweon
H