Heterologous Expression of Histidine Acid Phytase from Pantoea sp. 3.5.1 in Yarrowia lipolytica
- 19 Downloads
The dimorphic yeasts Yarrowia lipolytica are used as effective expression system and are characterized by a high level of production of heterologous protein. In this work, we aimed to clone and express Pantoea sp. 3.5.1 agpP phytase gene in Yarrowia lipolytica. Genetic constructs containing the native phytase gene (agpP) under the control of the strong hybrid promoter hp4d and the signal peptide of the alkaline extracellular protease XPR2 gene, as well as the optimized gene (agpP-opt) of phytase under the control of two signal peptides—bacterial and yeast were obtained. Recombinant Y. lipolytica strains with integrated bacterial phytase genes were obtained and expression of phytase was analyzed.
KeywordsPhytase Cloning Heterologous expression Yarrowia
This work was performed in accordance with the Russian Government Program of Competitive Growth of the Kazan Federal University and supported by the Russian Foundation for Basic Research (project no. 16-34-60191).
- 1.Suzuki, U., Yoshimura, K., & Takaishi, M. (1907). Über ein enzym ‘Phytase’ das anhydro-oxy-methylen diphosphorsaure’ spalter. Bulletin of the College of Agriculture, Tokyo Imperial University, 7, 503–512.Google Scholar
- 2.Hayakawa, T., Suzuki, K., Miura, H., Ohno, T., & Igaue, I. (1990). Myo-inositol polyphosphate intermediates in the dephosphorylation of Phytic acid by acid phosphatase with phytase activity from rice bran. Agricultural and Biological Chemistry, 54(2), 279–286.Google Scholar
- 3.Thomas, M. P., Mills, S. J., & Potter, B. V. L. (2016). The “other” Inositols and their phosphates: synthesis, biology, and medicine (with recent advances in myoInositol chemistry). Angewandte Chemie, International Edition, 55, 1614–1650. https://doi.org/10.1002/anie.201502227.CrossRefGoogle Scholar
- 7.Siren M. (1986a). Stabilized pharmaceutical and biological material composition. Pat. SE 003165.Google Scholar
- 8.Siren M. (1986b). New myo-inositol triphosphoric acid isomer. Pat.SW 052950.Google Scholar
- 9.Suleimanova, A. D., Beinhauer, A., Valeeva, L. R., Chastukhina, I. B., Balaban, N. P., Shakirov, E. V., Greiner, R., & Sharipova, M. R. (2015). Novel glucose-1-phosphatase with high phytase activity and unusual metal ion activation from soil bacterium Pantoea sp. strain 3.5.1. Applied and Environmental Microbiology, 81, 6790–6799. https://doi.org/10.1128/AEM.01384-15.CrossRefGoogle Scholar
- 12.Madzak, C., Treton, B., & Blanchin-Roland, S. (2000). Strong hybrid promoters and integrative expression/secretion vectors for quasiconstitutive expression of heterologous proteins in the yeast Yarrowia lipolytica. Journal of Molecular Microbiology and Biotechnology, 2, 207–216.Google Scholar
- 13.Sambrook, J., & Russell, D. W. (2001). Molecular cloning: a laboratory manual. Cold: Cold Spring Harbor Laboratory Press.Google Scholar