Expression of bacterial hemoglobin in the yeast, Pichia pastoris, with a low O2-induced promoter

• Published in: Biotechnology letters Vol. 27; no. 19; pp. 1491 - 1497
• Main Authors: CHIEN, Liang-Jung, LEE, Cheng-Kang
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.10.2005; Springer Nature B.V
• Subjects: Alcohol Dehydrogenase - genetics; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biological and medical sciences; Biotechnology; Blotting, Northern; Cloning, Molecular; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Fungal; Gene Transfer Techniques; Hemoglobins - genetics; Hemoglobins - metabolism; Microbiology; Oxygen - pharmacology; Pichia - enzymology; Pichia - genetics; Plasmids; Promoter Regions, Genetic; Protein Engineering; Recombinant Proteins; Truncated Hemoglobins; Yeasts; Oxygen; Yeast; Enzyme; Alcohol dehydrogenase; Pichia pastoris; Pichia; Fungi; Regulation(control); Hemoglobin; Bacteria; Ascomycetes; bacterial hemoglobin; Oxidoreductases; ADH2-promoter; Thallophyta; oxygen-regulated expression
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-005-1324-x

The PsADH2-promoter of Pichia stipitis alcohol dehydrogenase II (ADH II) gene was employed to control the expression of Vitreoscilla hemoglobin (VHb) gene in Pichia pastoris. As in P. stipitis, the promoter was also induced microaerobically in P. pastoris. The expression level of VHb in P. pastoris at low O2 tension (<5% air saturation) was 16 nmol/g dry cell wt, i.e. about 24-fold higher than that at 60% air saturation. The expressed VHb enhanced growth of P. pastoris under microaerobic conditions. The application of O2-regulated promoter in P. pastoris revealed that induction of high-level expression of heterologous protein is feasible without addition of supplementary compounds.