Effects of methanol feeding methods on chimeric α-amylase expression in continuous culture of Pichia pastoris

2006 ◽  
Vol 101 (3) ◽  
pp. 227-231 ◽  
Author(s):  
Atsushi Nakano ◽  
Chun Yeon Lee ◽  
Arei Yoshida ◽  
Takehiro Matsumoto ◽  
Naofumi Shiomi ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Continuous Culture ◽  
Feeding Methods ◽  
Methanol Feeding

A new methanol-feeding strategy for the improved production of β-galactosidase in high cell-density fed-batch cultures of Pichia pastoris Mut+ strains

2012 ◽  
Vol 17 (1) ◽  
pp. 76-83 ◽  
Author(s):  
Amir Maghsoudi ◽  
Safoura Hosseini ◽  
Seyed Abbas Shojaosadati ◽  
Ebrahim Vasheghani-Farahani ◽  
Mohsen Nosrati ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Cell Density ◽  
High Cell Density ◽  
Feeding Strategy ◽  
High Cell ◽  
Fed Batch ◽  
Batch Cultures ◽  
Methanol Feeding

scholarly journals Effect of Methanol Feeding Rate on the Expression of Glomerella cingulata Cutinase Using Recombinant Pichia pastoris in Shake Flask

2014 ◽  
Vol 14 (13) ◽  
pp. 1430-1434
Author(s):  
Rahimi Kamarud-Di ◽  
Jamaliah Md. Jahim ◽  
Farah Diba Abu B ◽  
Wahab Mohamed ◽  
Osman Hassan
Keyword(s):  
Pichia Pastoris ◽  
Shake Flask ◽  
Feeding Rate ◽  
Glomerella Cingulata ◽  
Recombinant Pichia Pastoris ◽  
Methanol Feeding

scholarly journals Controlling methanol feeding for recombinant protein production by Pichia pastoris under oxidation stress in fed-batch fermentation

2020 ◽  
Author(s):  
Rongkang Hu ◽  
Dongming Lan ◽  
Ruiguo Cui ◽  
Haomin Hong ◽  
Yachun Niu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Pichia Pastoris ◽  
Batch Fermentation ◽  
Protein Production ◽  
Feeding Rate ◽  
Induction Phase ◽  
Fed Batch ◽  
Ros Accumulation ◽  
Fed Batch Fermentation ◽  
Methanol Feeding

Abstract Background: Methanol can be used by Pichia pastoris as the sole carbon source and inducer to produce recombinant proteins in high-cell-density fermentations, but also damages cells due to reactive oxygen species (ROS) accumulation from methanol oxidation. Here, we study the relationship between methanol feeding and ROS accumulation by controlling constant methanol feeding rate during the induction phase.Results: Higher methanol feeding rate increased the level of ROS accumulation caused by methanol oxidation. While the cell growth rate was proportional to the rate of methanol feeding rate, but maximum total protein production and highest enzyme activity were achieved at methanol feeding rate 4 mL/(L·h) as compared to that with 5 mL/(L·h). Moreover, oxidative demage induced by over accumulation of ROS in P. pastoris during the methanol induction phase caused cell death and reduced protein expression ability. ROS scavenging system analysis reveals that the higher methanol feeding rate, especially 5 mL/(L·h), resulted in increased intracellular catalase activity and decreased glutathione content significantly. Finally, Spearman's correlation analysis further reveals that the reduced glutathione might be beneficial for maintaining cell viability and increasing protein production under oxidative stress caused by toxic accumulation of ROS accumulation.Conclusion: Our findings suggest an integrated strategy to control the feeding of the essential substrate based on analyzing its response to oxidative stress caused by toxic accumulation of ROS accumulation, as well as develop strategy to optimize fed-batch fermentation.

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