Impacts of Magnetic Immobilization on the Growth and Metabolic Status of Recombinant Pichia pastoris

Mol Biotechnol. 2022 Mar;64(3):320-329. doi: 10.1007/s12033-021-00420-w. Epub 2021 Oct 13.

Abstract

Downstream processing is an expensive step for industrial production of recombinant proteins. Cell immobilization is known as one of the ideal solutions in regard to process intensification. In recent years, magnetic immobilization was introduced as a new technique for cell immobilization. This technique was successfully employed to harvest many bacterial and eukaryotic cells. But there are no data about the influence of magnetic immobilization on the eukaryotic inducted recombinant cells. In this study, impacts of magnetic immobilization on the growth and metabolic status of induced recombinant Pichia pastoris as a valuable eukaryotic model cells were investigated. Results based on colony-forming unit, OD600, and trypan blue assay indicated that magnetic immobilization had no adverse effect on the growth and viability of P. pastoris cells. Also, about 20-40% increase in metabolic activity was recorded in immobilized cells that were decorated with 0.5-2 mg/mL nanoparticles. Total protein and carbohydrate of the cells were also measured as main indicatives for cell function and no significant changes were observed in the immobilized cells. Current data show magnetic immobilization as a biocompatible technique for application in eukaryotic expression systems. Results can be considered for further developments in P. pastoris-based expression systems.

Keywords: Magnetic nanoparticles; Magnetic separation; Metabolism; Recombinant protein; Yeast cells.

MeSH terms

  • Magnetic Phenomena
  • Magnetite Nanoparticles / chemistry*
  • Microbial Viability
  • Microbiological Techniques / methods*
  • Pichia / genetics
  • Pichia / growth & development*
  • Recombination, Genetic

Substances

  • Magnetite Nanoparticles