Enhanced porcine circovirus Cap protein production by Pichia pastoris with a fuzzy logic DO control based methanol/sorbitol co-feeding induction strategy

Ding, Jian; Zhang, Chunyu; Gao, Minjie; Hou, Guoli; Liang, Kexue; Li, Chunhua; Ni, Jianping; Li, Zhen; Shi, Zhongping

doi:10.1016/j.jbiotec.2014.02.018

Cited by 12 publications

(3 citation statements)

References 31 publications

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“…Thus, to use carbon sources that support growth but do not repress the AOX1 promoter is essential to control and scale-up fermentation process. On the other hand, sorbitol is a widely accepted non-repressive carbon source for P. pastoris, which can produce a higher level of foreign protein compared with glycerol [29,30]. Arnau et al [31] made a systematic study of the influence of methanol setpoint and sorbitol feeding rate in fed-batch operation with a P. pastoris strain producing Rhizopus oryzae lipase (ROL), and ROL production and Y P/X were 1.25 fold higher and volumetric and specific productivity were 1.35 fold higher using mixed substrates comparing with the methanol as sole carbon source.…”

Section: Discussionmentioning

confidence: 99%

Enhanced protein production by sorbitol co-feeding with methanol in recombinant Pichia pastoris strains

Chen

Mohsin

Chu

et al. 2017

Biotechnol Bioproc E

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Pichia pastoris strains carrying 1, 6, 12, and 18 copies of the porcine insulin precursor (PIP) gene, were employed to investigate the effects of sorbitol co-feeding with methanol on the physiology of the strains. Multicopy clones of the methylotrophic yeast were generated to vary the PIP gene dosage and recombinant proteins. Elevated gene dosage increased levels of the recombinant PIP protein when methanol served as the sole carbon and energy source i.e., an increase of 1.9% for a strain carrying 1 copy, 42.6% for a strain carrying 6 copies, 34.7% for a strain carrying 12 copies and 80.9% for a strain carrying 18 copies, respectively (using sorbitol co-feeding with methanol during the induction phase). However, it had no significant influence on a lower gene dosage strain (1 copy), but this approach affirmed enhancement in cell growth and PIP production for higher gene dosage strain (6, 12, and 18 copies) via using sorbitol co-feeding with methanol. Additionally, the co-feeding strategy could hold vital importance for recombinant protein production by a multi-copy P. pastoris system.

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Section: Discussionmentioning

confidence: 99%

Enhanced protein production by sorbitol co-feeding with methanol in recombinant Pichia pastoris strains

Chen

Mohsin

Chu

et al. 2017

Biotechnol Bioproc E

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“…It was reported that once the concentration of methanol was kept at a constant, the expression level of foreign protein could reach the highest . Also, the mixed feeding strategy was commonly used recent years, such as glycerol–methanol methanol–sorbitol cofeeding , which would be the next striving orientation. After optimization of the induction pH, induction T , and V m , the maximum production of HSA/IL1Ra was about 1.1 g/L under the final optimized culture condition (pH 7.0, T 29 ℃, and V m 4.82 mL/L/H).…”

Section: Discussionmentioning

confidence: 99%

RSM optimization of HSA/IL1Ra in Pichia pastoris overexpression strain and study of its in vivo activity in reducing hyperglycemia of GK rats

Liu

Zhao

Lai

et al. 2017

Biotech and App Biochem

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Human serum albumin (HSA) and interleukin-1 receptor antagonist (IL1Ra) fusion protein is a potential long-acting drug in the treatment of type 2 diabetes. Previously, the expression level of HSA/IL1Ra in Pichia pastoris was successfully improved by increasing the gene copy number and coexpression with chaperone (protein disulfide isomerase) in our laboratory. However, the overexpression strain resulted in low production of high- cell-density fermentation. In this study, the culture medium was optimized in both flask and fermenter, and the optimum culture medium notably increased the productivity and stability of HSA/IL1Ra. To further improve the expression, response surface methodology was used to further optimize the culture condition through modeling three selected parameters (induction pH, induction temperature [T], and maximum methanol feed rate [V ]). The maximum yield of HSA/IL1Ra reached 1.1 g/L (10-fold higher than original fermentation condition) under the optimized culture condition (pH 7.0, T = 29 ℃ and V = 4.82 mL/L/H) in a 5-L fermenter. In addition, the degradation position of HSA/IL1Ra during fermentation was determined to be K571, serving as a potential target for genetic modification strategies to reduce the degradation. Finally, the in vivo activity study showed that HSA/IL1Ra maintained the therapeutic effect of IL1Ra in type 2 diabetes model rats meanwhile reducing the frequency of administration.

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“…The time profile of cell concentrations and DO, as well as stirrer speed were depicted in Figure 7. Heterologous protein production by fed-batch culture with P. pastoris is mainly divided into two phases: a growth phase to accumulate large amount of cells with glycerol as the carbon source, and an induction phase by feeding methanol for heterologous protein expression [12,27]. 7.…”

Section: Performance During P Pastoris Fermentationmentioning

confidence: 99%

On-Line Tendency Control of Dissolved Oxygen Concentration during Aerobic Fed-Batch Fermentations

Zheng

Pan

2019

Applied Sciences

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In this paper, an on-line control strategy that aims to guarantee the maintenance of a minimum dissolved oxygen (DO) concentration during aerobic fed-batch fermentations is proposed. It is a difficult task to maintain the DO concentration, especially during fed-batch fermentation, due to strongly nonlinear, variable conditions and probe dynamics. The algorithm uses information contained in the slope of the profile of the DO, as this evolves in a timely way to adapt to process variations. Moving window technology was used to track the DO tendency variation. This method was tested in Corynebacterium glutamicum and Pichia pastoris fermentations. The performance of tendency control was compared with that of manual control. The experimental results clearly show that the on-line tendency control of DO is effective and can also reduce the frequency of activity of the controller as well as the manpower burden.

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Enhanced porcine circovirus Cap protein production by Pichia pastoris with a fuzzy logic DO control based methanol/sorbitol co-feeding induction strategy

Cited by 12 publications

References 31 publications

Enhanced protein production by sorbitol co-feeding with methanol in recombinant Pichia pastoris strains

Enhanced protein production by sorbitol co-feeding with methanol in recombinant Pichia pastoris strains

RSM optimization of HSA/IL1Ra in Pichia pastoris overexpression strain and study of its in vivo activity in reducing hyperglycemia of GK rats

On-Line Tendency Control of Dissolved Oxygen Concentration during Aerobic Fed-Batch Fermentations

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