Volume 25, Issue 4 (7-2021)                   IBJ 2021, 25(4): 255-264 | Back to browse issues page

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Zarei N, Ghasemi H, Nayebhashemi M, Zahmatkesh M, Jamalkhah M, Moeinian N, et al . Targeted Deletion of Los1 Homologue Affects the Production of a Recombinant Model Protein in Pichia pastoris. IBJ 2021; 25 (4) :255-264
URL: http://ibj.pasteur.ac.ir/article-1-3460-en.html
Targeted Deletion of Los1 Homologue Affects the Production of a Recombinant Model Protein in Pichia pastoris
Najmeh Zarei , Hosnieh Ghasemi , Mahsa Nayebhashemi , Mozhgan Zahmatkesh , Monire Jamalkhah , Nafiseh Moeinian , Zahra Mohammadi , Somayeh Enayati , Vahid Khalaj *
Abstract:  
Background: The methylotrophic yeast Pichia pastoris is an appealing production host for a variety of recombinant proteins, including biologics. In this sense, various genetic- and non-genetic-based techniques have been implemented to improve the production efficiency of this expression platform. Loss of supression (Los1) encodes a non-essential nuclear tRNA exporter in Saccharomyces cerevisiae, which its deletion extends replicative lifespan. Herein, a los1-deficient strain of P. pastoris was generated and characterized. Methods: A gene disruption cassette was prepared and transformed into an anti-CD22-expressing strain of P. pastoris. A δ los1 mutant was isolated and confirmed. The drug sensitivity of the mutant was also assessed. The growth pattern and the level of anti-CD22 single-chain variable fragment (scFv) expression were compared between the parent and mutant strains. Results: The los1 homologue was found to be a non-essential gene in P. pastoris. Furthermore, the susceptibility of los1 deletion strain to protein synthesis inhibitors was altered. This strain showed an approximately 1.85-fold increase in the extracellular level of anti-CD22 scFv (p < 0.05). The maximum concentrations of total proteins secreted by δ los1 and parent strains were 125 mg/L and 68 mg/L, respectively. Conclusion: The presented data suggest that the targeted disruption of los1 homologue in P. pastoris can result in a higher expression level of our target protein. Findings of this study may improve the current strategies used in optimizing the productivity of recombinant P. pastoris strains.
Keywords: Aging, Longevity, Pichia pastoris, Recombinant proteins
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Type of Study: Full Length/Original Article | Subject: Related Fields
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