Development of a strain of Hansenula polymorpha for the efficient expression of guar alpha-galactosidase

A strain of the methylotrophic yeast Hansenula polymorpha, A16, has been developed that expresses the guar alpha-galactosidase gene to 22.4 mg/g dry cell weight in chemostat cultures at a dilution rate of 0.1 h(-1). This corresponds to more than 13.1% of soluble cell protein, of which 56-62% is secr...

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Bibliographic Details
Published inYeast (Chichester, England) Vol. 8; no. 5; p. 361
Main Authors Veale, R A, Giuseppin, M L, van Eijk, H M, Sudbery, P E, Verrips, C T
Format Journal Article
LanguageEnglish
Published England 01.05.1992
Subjects
alpha-Galactosidase - biosynthesis
alpha-Galactosidase - genetics
Base Sequence
Biotechnology
DNA Probes
DNA, Fungal - genetics
Gene Expression
Molecular Sequence Data
Pichia - genetics
Plants - enzymology
Plants - genetics
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Summary:A strain of the methylotrophic yeast Hansenula polymorpha, A16, has been developed that expresses the guar alpha-galactosidase gene to 22.4 mg/g dry cell weight in chemostat cultures at a dilution rate of 0.1 h(-1). This corresponds to more than 13.1% of soluble cell protein, of which 56-62% is secreted into the medium. The alpha-galactosidase gene was flanked by the promoter and terminator sequences of the H.polymorpha mox gene, which can direct expression of the mox gene itself more than 30% of total cell protein under methanol growth. The expression cassette (pUR3510) based on the Saccharomyces cerevisiae plasmid, YEp13, was integrated into the genome. Such transformants were stable in chemostat cultures and exhibited 100% stability for both alpha-galactosidase+ and leu+ phenotypes. Chemostat cultures produced higher levels of alpha-galactosidase with higher specific productivities expressed as mg alpha-galactosidase g(-1) h(-1) compared to batch cultures.
ISSN:0749-503X
DOI:10.1002/yea.320080504