Optimization of Culture Parameters for Expression of Recombinant Human Basic Fibroblast Growth Factor in <i>Pichia pastoris</i> 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.

Oil-degrading bacterial consortium from Gulf of Mexico designed by a factorial method, reveals stable population dynamics

“…One of these methods is the Plackett-Burman (PB) design, which enables the identification of dominant factors in a system through the analysis of their orthogonal arrangements (Box et al, 2005;Lv et al, 2014). Applications of the PB method include the optimization of bacterial culture growth parameters (Jiang et al, 2013), design of microbial culture media (Prajapati et al, 2014;Ghanem and Al-Zahrani, 2016), and setting of parameters that influence metal removal by bacteria (Migahed et al, 2017). In the design of bacterial consortia, PB has been used to improve a synthetic bacterial consortium to treat dairy industry wastewater (Mazzucotelli et al, 2014).…”

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

“…Statistical methods for experimental design such as response surface methodology (RSM) have been used to optimize the key variables during microbiological fermentation and achieve the expected outcomes . In the present study, the effects of various fermentation parameters on the expression of HSA/IL1Ra in P. pastoris were discussed, such as induction T , induction pH, dissolved oxygen (DO), V m , and nitrogen source supplementation (YNB, yeast extract, and peptone).…”