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Constitutive and extracellular expression of pectin methylesterase from Pectobacterium chrysanthemi in Pichia pastoris

Abstract

Pectin methylesterase (PME) which is widely used in the cosmetic, food and pharmaceutical industries catalyses the hydrolysis of the methyl ester of pectin to yield methanol and free carboxyl groups. This study was performed to produce active pectin methylesterase (PME) extracellularly from Pectobacterium chrysanthemi in Pichia pastoris. Firstly, pGKBα was constructed for the secretion of heterologous protein. After it was cloned in Escherichia coli cells and the sequence was affirmed, PME gene was inserted into pGKBα. So, pGKBα-PME carried the PME gene in correct position was cloned in E. coli cells. Then, P. pastoris X-33 cells were transformed with linearized pGKBα-PME and six transformants were cultivated for recombinant PME production. It was observed that one of them had a high-capacity secretion of active PME. The molecular mass of extracellular PME enzyme was found to be about 59 kDa. The PME enzyme from P. chrysanthemi was produced by P. pastoris for the first time in this study. This recombinant enzyme might be produced in a large scale and also purified from the culture medium. Then, the purified enzyme might be used for clarification and increasing yield of juice in food industrial applications.

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Acknowledgements

This work was supported by the Scientific Research Foundation of Atatürk University (FYL2018-6853). The authors would like to acknowledge East Anatolia High Technology Application and Research Center (DAYTAM, Erzurum, Turkey) for the technical support.

Funding

This work was supported by the Scientific Research Foundation of Ataturk University (FYL-2018–6853).

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Correspondence to Yagmur Unver.

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Acar, M., Unver, Y. Constitutive and extracellular expression of pectin methylesterase from Pectobacterium chrysanthemi in Pichia pastoris. 3 Biotech 12, 219 (2022). https://doi.org/10.1007/s13205-022-03291-3

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  • DOI: https://doi.org/10.1007/s13205-022-03291-3

Keywords

  • Pectin methylesterase
  • Expression
  • Pichia pastoris
  • pGKBα
  • Pectobacterium chrysanthemi