Recombinant Hansenula polymorpha as a biocatalyst: coexpression of the spinach glycolate oxidase (GO) and the S. cerevisiae catalase T (CTT1) gene

• Published in: Applied microbiology and biotechnology Vol. 46; no. 1; pp. 46 - 54
• Main Authors: Gellissen, G, Piontek, M, Dahlems, U, Jenzelewski, V, Gavagan, J.E, DiCosimo, R, Anton, D.L, Janowicz, Z.A
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01-08-1996
• Subjects: Alcohol Oxidoreductases - genetics; Alcohol Oxidoreductases - metabolism; Biological and medical sciences; Biological Sciences; Biotechnology; Catalase - genetics; Catalase - metabolism; Catalysis; Cytosol - enzymology; Fermentation; Fundamental and applied biological sciences. Psychology; Gene Expression; Genetic engineering; Genetic technics; Glycolates - metabolism; Glyoxylates - metabolism; Industrial Microbiology; Methods. Procedures. Technologies; Microbodies - enzymology; Modification of gene expression level; Pichia - enzymology; Pichia - genetics; Recombinant Proteins - biosynthesis; Saccharomyces cerevisiae - enzymology; Spinacia oleracea - enzymology; Recombinant microorganism; Yeast; Coexpression; Spinacia oleracea; Glyoxylic acid; Fungi; Catalase; Dicotyledones; Biotransformation; Glycolic acid; Angiospermae; (S)-2-Hydroxy-acid oxidase; Ascomycetes; Recombinant protein; Thallophyta; Heterologous system; Enzyme; Gene expression; Chenopodiaceae; Peroxidases; Microorganism culture; Spermatophyta; Oxidoreductases; Application; Hansenula polymorpha; Saccharomyces cerevisiae
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s002530050781

The methylotrophic yeast Hansenula polymorpha has been developed as an efficient production system for heterologous proteins. The system offers the possibility to cointegrate heterologous genes in anticipated fixed copy numbers into the chromosome. As a consequence coproduction of different proteins in stoichiometric ratios can be envisaged. This provides options to design this yeast as an industrial biocatalyst in procedures where several enzymes are required for the efficient conversion of a given inexpensive compound into a valuable product. To this end recombinant strains have been engineered with multiple copies of expression cassettes containing the glycolate oxidase (GO) gene from spinach and the catalase T (CTT1 ) gene from S. cerevisiae. The newly created strains produce high levels of the peroxisomal glycolate oxidase and the cytosolic catalase T. The strains efficiently convert glycolate into glyoxylic acid, oxidizing the added substrate and decomposing the peroxide formed during this reaction into water and oxygen.