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Applied and Environmental Microbiology, June 2009, p. 3528-3534, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.02913-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Phosphate-Responsive Promoter of a Pichia pastoris Sodium Phosphate Symporter{triangledown} ,{dagger}

Jungoh Ahn,1 Jiyeon Hong,1 Myongsoo Park,1 Hyeokweon Lee,1 Eungyo Lee,1 Chunsuk Kim,1 Joohwan Lee,1 Eui-sung Choi,2 Joon-ki Jung,1 and Hongweon Lee1*

Biotechnology Process Engineering Center,1 Systems Microbiology Research Center, KRIBB, Daejeon 305-806, Republic of Korea2

Received 22 December 2008/ Accepted 20 March 2009

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 1{alpha} 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.

* Corresponding author. Mailing address: Biotechnology Process Engineering Center, KRIBB, Daejeon 305-600, Republic of Korea. Phone: 82-42-860-4520. Fax: 82-42-860-4516. E-mail: hwlee{at}kribb.re.kr

{triangledown} Published ahead of print on 27 March 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, June 2009, p. 3528-3534, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.02913-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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