Affiliation:
1. Biotechnology Process Engineering Center
2. Systems Microbiology Research Center, KRIBB, Daejeon 305-806, Republic of Korea
Abstract
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 1α 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
.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
41 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献