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Increasement of Heterologous Expression of Recombinant Vit v 1 in Pichia pastoris KM71 by Nonionic Detergents as a Cost-effective Approach

Abstract

Vit v 1 as a lipid-transfer protein is a major allergen of grapes (Vitis vinifera) that elicits food allergy in many patients in Iran. Todays, recombinant allergens have a promising future in improving in vivo and in vitro diagnostic tests. In this study, we aim to achieve high-level expression of recombinant Vit v 1 allergen in Pichia pastoris expression system. DNA construct has been cloned into the pGAPZαC plasmid and expressed in the methylotrophic yeast P. pastoris KM71. The maximum yield obtained after 4 days of expression under the optimal conditions (at 25°C with 0.05% Triton X-100). The secreted recombinant Vit v 1 was purified by nickel-affinity chromatography. The results showed that the recombinant Vit v 1 was expressed mainly as the soluble protein with an apparent molecular mass of 37.8 kDa in the SDS-PAGE and Western blotting. The findings suggest that nonionic detergents could increase yield of recombinant Vit v 1, probably through enhancing transfer of oxygen and membrane lipids fluidity.

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Funding

We acknowledge the funding support from Research Administration Department of Mashhad University of Medical Sciences (Grant no: 88800).

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Correspondence to M. Sankian.

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The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Moghadam, M., Ayati, H., Shobeiri, S. et al. Increasement of Heterologous Expression of Recombinant Vit v 1 in Pichia pastoris KM71 by Nonionic Detergents as a Cost-effective Approach. Appl Biochem Microbiol 58, 443–448 (2022). https://doi.org/10.1134/S0003683822040111

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  • DOI: https://doi.org/10.1134/S0003683822040111

Keywords:

  • grape allergen
  • Vit v 1
  • Pichia pastoris
  • KM71
  • LTP
  • recombinant allergen