Development of a dual specific growth rate-based fed-batch process for production of recombinant human granulocyte colony-stimulating factor in Pichia pastoris

2020 ◽  
Author(s):  
Ashwani Gautam ◽  
Vikram Sahai ◽  
Saroj Mishra
Keyword(s):  
Growth Rate ◽  
Pichia Pastoris ◽  
Specific Growth Rate ◽  
Specific Growth ◽  
Batch Process ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Fed Batch ◽  
Stimulating Factor

scholarly journals Specific growth rate governs AOX1 gene expression, affecting the production kinetics of Pichia pastoris (Komagataella phaffii) PAOX1-driven recombinant producer strains with different target gene dosage

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Javier Garrigós-Martínez ◽  
Miguel Angel Nieto-Taype ◽  
Arnau Gasset-Franch ◽  
José Luis Montesinos-Seguí ◽  
Xavier Garcia-Ortega ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Rate ◽  
Pichia Pastoris ◽  
Specific Growth Rate ◽  
Specific Growth ◽  
Gene Dosage ◽  
Fed Batch ◽  
Komagataella Phaffii ◽  
Production Kinetics ◽  
Kinetics Of

Abstract Background The PAOX1-based expression system is the most widely used for producing recombinant proteins in the methylotrophic yeast Pichia pastoris (Komagataella phaffii). Despite relevant recent advances in regulation of the methanol utilization (MUT) pathway have been made, the role of specific growth rate (µ) in AOX1 regulation remains unknown, and therefore, its impact on protein production kinetics is still unclear. Results The influence of heterologous gene dosage, and both, operational mode and strategy, on culture physiological state was studied by cultivating the two PAOX1-driven Candida rugosa lipase 1 (Crl1) producer clones. Specifically, a clone integrating a single expression cassette of CRL1 was compared with one containing three cassettes over broad dilution rate and µ ranges in both chemostat and fed-batch cultivations. Chemostat cultivations allowed to establish the impact of µ on the MUT-related MIT1 pool which leads to a bell-shaped relationship between µ and PAOX1-driven gene expression, influencing directly Crl1 production kinetics. Also, chemostat and fed-batch cultivations exposed the favorable effects of increasing the CRL1 gene dosage (up to 2.4 fold in qp) on Crl1 production with no significant detrimental effects on physiological capabilities. Conclusions PAOX1-driven gene expression and Crl1 production kinetics in P. pastoris were successfully correlated with µ. In fact, µ governs MUT-related MIT1 amount that triggers PAOX1-driven gene expression—heterologous genes included—, thus directly influencing the production kinetics of recombinant protein.

scholarly journals Fed-batch fermentation of GM-CSF-producing glycoengineered Pichia pastoris under controlled specific growth rate

2010 ◽  
Vol 9 (1) ◽  
pp. 93 ◽  
Author(s):  
Pieter P Jacobs ◽  
Mehmet Inan ◽  
Nele Festjens ◽  
Jurgen Haustraete ◽  
Annelies Van Hecke ◽  
...  
Keyword(s):  
Growth Rate ◽  
Pichia Pastoris ◽  
Specific Growth Rate ◽  
Batch Fermentation ◽  
Specific Growth ◽  
Fed Batch ◽  
Gm Csf ◽  
Fed Batch Fermentation ◽  
Controlled Specific Growth Rate

Over expression and analysis of O-glycosylated recombinant human granulocyte colony stimulating factor in Pichia pastoris using Agilent 2100 Bioanalyzer

2009 ◽  
Vol 143 (1) ◽  
pp. 44-50 ◽  
Author(s):  
Anjali Apte-Deshpande ◽  
Sandeep Somani ◽  
Goutam Mandal ◽  
Sudheerbabu Soorapaneni ◽  
Sriram Padmanabhan
Keyword(s):  
Pichia Pastoris ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Over Expression ◽  
Stimulating Factor

Influence of specific growth rate over the secretory expression of recombinant potato carboxypeptidase inhibitor in fed-batch cultures of Escherichia coli

2010 ◽  
Vol 45 (8) ◽  
pp. 1334-1341 ◽  
Author(s):  
Juan-Miguel Puertas ◽  
Jordi Ruiz ◽  
Mónica Rodríguez de la Vega ◽  
Julia Lorenzo ◽  
Glòria Caminal ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Growth Rate ◽  
Specific Growth Rate ◽  
Specific Growth ◽  
Fed Batch ◽  
Secretory Expression ◽  
Batch Cultures ◽  
Carboxypeptidase Inhibitor

An algorithm for operating a fed-batch fermentor at optimum specific-growth rate

1989 ◽  
Vol 33 (1) ◽  
pp. 115-125 ◽  
Author(s):  
Pramod Agrawal ◽  
George Koshy ◽  
Michael Ramseier
Keyword(s):  
Growth Rate ◽  
Specific Growth Rate ◽  
Specific Growth ◽  
Fed Batch
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