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Heterologous extracellular expression and initial characterization of the peroxisomal catalase from the methylotrophic yeast Hansenula polymorpha in Pichia pastoris

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Abstract

Catalase is well known to eliminate H2O2 in cells and reduces the toxicity of peroxide compounds. A catalase gene HpCAT1 of methylotrophic yeast Hansenula polymorpha without the part coding the native signal peptide was cloned into expression vector pYM3165 and then integrated into genome of Pichia pastoris GS115 by electroporation. The result of the enzyme activity assay and SDS-PAGE demonstrated that the recombinant protein (HpCAT1) of H. polymorpha was extracellularly expressed in P. pastoris. The expressed catalase was recovered from the culture supernatant of P. pastoris GS115 and purified by (NH4)2SO4 fractionation and Ni-NTA affinity chromatography. The main biochemical properties of the recombinant protein HpCAT1, such as thermodependence and thermostability, pH optimum and pH stability, as well as the effect of metal ions and chemicals, were characterized. With H2O2 as the substrate, HpCAT1 displayed pH and temperature optima of ∼2.6 and 45°C, respectively. The recombinant HpCAT1 activity was inhibited by 1 mM Hg2+ and Cu2+, but was highly enhanced by 1.0 mM Fe2+.

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Authors and Affiliations

  1. Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China

    Y. -S. Tian, H. Xu, J. Xu, R. -H. Peng & Q. -H. Yao

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  1. Y. -S. Tian
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  2. H. Xu
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  3. J. Xu
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  4. R. -H. Peng
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  5. Q. -H. Yao
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Correspondence to Q. -H. Yao.

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Original Russian Text © Y.-S. Tian, H. Xu, J. Xu, R.-H. Peng, Q.-H. Yao, 2013, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2013, Vol. 49, No. 5, pp. 504–512.

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Tian, Y.S., Xu, H., Xu, J. et al. Heterologous extracellular expression and initial characterization of the peroxisomal catalase from the methylotrophic yeast Hansenula polymorpha in Pichia pastoris . Appl Biochem Microbiol 49, 507–513 (2013). https://doi.org/10.1134/S0003683813050141

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