Engineering folding mechanism through Hsp70 and Hsp40 chaperones for enhancing the production of recombinant human interferon gamma (rhIFN-γ) in Pichia pastoris cell factory

Effect of gene copy number and chaperone coexpression on recombinant hydrophobin HFBI biosurfactant production in Pichia pastoris

“…HFBI expression alone caused a significant decrease in the growth rate and maximum OD 600 reached in the case of every HFBI copy number strain examined as compared with the GS115 parent strain growth curve (Figure ). Coexpression of Kar2p, Pdi1p, or Ero1p caused increased growth compared with the no chaperone strain in each copy number group, which is consistent with previous reports where coexpression of chaperones rescued yeast growth from the cytotoxic effects of target gene expression alone (Prabhu et al, ; W. Zhang et al, ). Thus, these results suggest expression of HFBI alone is inhibitory to normal cell growth, but overexpression of chaperones alleviates this effect, possibly by mitigating misfolding of the recombinant hydrophobin in the cell and thereby alleviating the stress on the cell.…”

“…To this end, recombinant protein production in P. pastoris has been greatly enhanced using metabolic engineering techniques such as overexpressing endogenous or exogenous chaperone proteins to promote recombinant protein folding and secretion (Damasceno et al, , ; Delic et al, ; Gasser, Maurer, Gach, Kunert, & Mattanovich, ; Gasser, Sauer, Maurer, Stadlmayr, & Mattanovich, ; Guerfal et al, ; Inan, Aryasomayajula, Sinha, & Meagher, ; Prabhu et al, ; Shen, Wu, Wang, Naranmandura, & Chen, ; Tsai, Duggan, Shimp Jr, Miller, & Narum, ; Yang et al, ; W. Zhang et al, ; J. Zhang, Wu, Chen, & Wu, ) and by increasing the target gene copy number (Aw & Polizzi, ; Clare et al, ; Scorer, Clare, McCombie, Romanos, & Sreekrishna, ; Shen et al, ; Yang et al, ). With regard to hydrophobins and their unique structure, molecular chaperones involved in preventing nonspecific hydrophobic aggregation and disulfide bond formation become obvious potential targets to increase recombinant expression.…”

Artificial neural network‐genetic algorithm (ANN‐GA) based medium optimization for the production of human interferon gamma (hIFN‐γ) in Kluyveromyces lactis cell factory

“…Unstructured kinetic models were used to simulate batch profiles with biomass concentration (X), substrate concentration (S), and hIFN‐γ concentration (P), in which the growth analysis was done using the logistic equation a substrate independent model that states that the “rate of the increase of cell biomass is proportional to the quantity of viable cell biomass at any instant time.” The logistic equation (Equation ) infers that growth of a cell is governed by a hyperbolic relation in which there is a limit to the maximum attainable biomass concentration . Equation is as follows: where dX/dt is the rate of biomass production (g/L/h), µ max is the maximum specific growth rate (h −1 ), X is the biomass concentration (g/L), and X max is the model predicted maximum biomass concentration (g/L).…”

“…However, the production level of hIFN‐γ in CHO cell lines were very less compared to E. coli , and, moreover, the CHO cell lines require a serum‐containing medium, which increases the production cost . Yeast expression system such as S. cerevisae and P. pastoris were also used for the heterologous production of hIFN‐γ. For these systems, being unicellular provides the advantage of easy and fastidious production and maintenance along with the provision of the required post‐translational modifications.…”

Accurate and cost‐effective prediction of HBsAg titer in industrial scale fermentation process of recombinant Pichia pastoris by using neural network based soft sensor

“…In the last four decades, implementation of recombinant DNA technology for manufacturing therapeutic proteins has been raised significantly . Among various host cells, the methylotrophic yeasts such as Pichia pastoris has gained major attention for large‐scale production as they cover somehow the advantages of both bacterial and mammalian cells . Recombinant P. pastoris can produce a high level of heterologous proteins in a simple and protein‐free medium, and it also performs post‐translation modification relatively similar to the mammalian cells .…”