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Molecular characterization of the Hansenula polymorpha FLD1 gene encoding formaldehyde dehydrogenase.Author(s): Richard J. Baerends; Grietje J. Sulter; Thomas W. Jeffries; James M. Cregg; Marten Veenhuis
Source: Yeast. Vol. 19 (2002): Pages 37-42.
Publication Series: Miscellaneous Publication
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DescriptionGlutathione-dependent formaldehyde dehydrogenase (FLD) is a key enzyme required for the catabolism of methanol as a carbon source and certain primary amines, such as methylamine as nitrogen sources in methylotrophic yeasts. Here we describe the molecular characterization of the FLD1 gene from the yeast Hansenula polymorpha. Unlike the recently described Pichia pastoris homologue, the H. polymorpha gene does not contain an intron. The predicted FLD1 product (Fld1p) is a protein of 380 amino acids (ca. 41 kDa) with 82% identity to P. pastoris Fld1p, 76% identity to the FLD protein sequence from n-alkane-assimilating yeast Candida maltosa and 63–64% identity to dehydrogenase class III enzymes from humans and other higher eukaryotes. The expression of FLD1 is strictly regulated and can be controlled at two expression levels by manipulation of the growth conditions. The gene is strongly induced under methylotrophic growth conditions; moderate expression is obtained under conditions in which a primary amine, e.g. methylamine, is used as nitrogen source. These properties render the FLD1 promoter of high interest for heterologous gene expression. The availability of the H. polymorpha FLD1 promoter provides an attractive alternative for expression of foreign genes besides the commonly used alcohol oxidase promoter. The sequence has been deposited in the GenBank/NCBI data library under Accession No. AF364077.
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CitationBaerends, Richard J.; Sulter, Grietje J.; Jeffries, Thomas W.; Cregg, James M.; Veenhuis, Marten. 2002. Molecular characterization of the Hansenula polymorpha FLD1 gene encoding formaldehyde dehydrogenase. Yeast. Vol. 19 (2002): Pages 37-42.
KeywordsHeterologous gene expression, methylotrophic yeast, methanol metabolism.
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