High cell density fermentation via a metabolically engineered Escherichia coli for the enhanced production of succinic acid

2011 ◽  
Vol 86 (4) ◽  
pp. 512-518 ◽  
Author(s):  
Dan Wang ◽  
Qiang Li ◽  
Ziyu Song ◽  
Wei Zhou ◽  
Zhiguo Su ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Succinic Acid ◽  
Cell Density ◽  
High Cell Density ◽  
High Cell ◽  
Enhanced Production ◽  
High Cell Density Fermentation ◽  
Engineered Escherichia Coli ◽  
Metabolically Engineered Escherichia Coli

scholarly journals Media optimization for SHuffle T7 Escherichia coli expressing SUMO-Lispro proinsulin by response surface methodology

2022 ◽  
Vol 22 (1) ◽  
Author(s):  
Aida Bakhshi Khalilvand ◽  
Saeed Aminzadeh ◽  
Mohammad Hossein Sanati ◽  
Fereidoun Mahboudi
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
Shake Flask ◽  
High Cell Density ◽  
Fold Increase ◽  
Soluble Expression ◽  
High Yield ◽  
High Cell ◽  
E Coli ◽  
High Cell Density Fermentation

Abstract Background SHuffle is a suitable Escherichia coli (E. coli) strain for high yield cytoplasmic soluble expression of disulfide-bonded proteins such as Insulin due to its oxidative cytoplasmic condition and the ability to correct the arrangement of disulfide bonds. Lispro is an Insulin analog that is conventionally produced in E. coli as inclusion bodies (IBs) with prolonged production time and low recovery. Here in this study, we aimed to optimize cultivation media composition for high cell density fermentation of SHuffle T7 E. coli expressing soluble Lispro proinsulin fused to SUMO tag (SU-INS construct) to obtain high cell density fermentation. Results Factors including carbon and nitrogen sources, salts, metal ions, and pH were screened via Plackett–Burman design for their effectiveness on cell dry weight (CDW) as a measure of cell growth. The most significant variables of the screening experiment were Yeast extract and MgCl2 concentration, as well as pH. Succeedingly, The Central Composite Design was utilized to further evaluate and optimize the level of significant variables. The Optimized media (OM-I) enhanced biomass by 2.3 fold in the shake flask (2.5 g/L CDW) that reached 6.45 g/L (2.6 fold increase) when applied in batch culture fermentation. The efficacy of OM-I media for soluble expression was confirmed in both shake flask and fermentor. Conclusion The proposed media was suitable for high cell density fermentation of E. coli SHuffle T7 and was applicable for high yield soluble expression of Lispro proinsulin.

Sign in / Sign up

Export Citation Format

Share Document