Optimization of Culture Parameters for Expression of Recombinant Human Basic Fibroblast Growth Factor in Pichia pastoris Using Response Surface Methodology Combining with Plackett-Burman Design

Rui Jiang; Li Wei Sun; Yu Zhao; Kai Feng; Shou Yan Chen

doi:10.4028/www.scientific.net/AMR.647.456

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Optimization of Culture Parameters for Expression of Recombinant Human Basic Fibroblast Growth Factor in Pichia pastoris Using Response Surface Methodology Combining with Plackett-Burman Design

Abstract:

In this study, we used plackett-burman design (PBD) and response surface methodology (RSM) methods to optimize media and culture conditions for expression of recombinant human basic fibroblast growth factor (rhbFGF) in Pichia pastoris in order to effectively enhance the production of rhbFGF. PBD was used to screen the most significant factors from nine important factors, which were fermentation time, methanol, temperature, initial pH, OD600 for inducement, concentrated volume, YNB peptone, and casein. Then RSM was further used to optimize the significant parameters. Our results had shown that three significant factors selected by PBD were temperature, fermentation time, and methanol. Base on the 3-factor-3-level central composite design (CCD) of RSM, the final optimized conditions were fermentation time 126.7h, methanol 1.3%, temperature 21.3°C, initial pH 6.0±0.1, OD600 for inducement 26, concentrated volume 100 ml, YNB 13.4%, peptone 4%, and casein 0.1%. The maximum production of rhbFGF was 302.4mg/L, which was 2 times higher than the production of rhbFGF reported in the previous studies. Those results provided the foundation for further researches on higher density cultivation condition of rhbFGF explored in the fermenter and accelerate the industrialization development of rhbFGF.