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- W2372233312 abstract "The fermentation conditions of the Escherichia coli strain [pET32a-Mtil-BL(DE3) 21] expressing a recombinant maleate cis-trans isomerase were optimized by orthogonal experiment design.The effects of the components of the culture medium(tryptone,yeast extract and pH),shaking speed,medium volume and inoculum concentration on cell growth and expression of maleate cis-trans isomerase were studied in 250 mL erlenmeyer flask containing 50 mL medium.The optimum medium composition was as follows: Tryptone 20 g L-1,yeast extract 10 g L-1,K2HPO4·3H2O 3.0 g L-1,KH2PO4 1.5 g L-1,NaCl 6 g L-1,MgSO4 3 g L-1,pH 6.5.The optimum conditions for cell growth and expression of maleate cis-trans isomerase were as follows: Inoculum concentration 5%,medium volume 20%,37 ℃ and shaking speed 220 r min-1.When D600 nm reached approximately 1.0,cells were induced by addition of 0.05 mmol L-1 of IPTG.Growth was continued at 37 ℃ for up to 6 h.The fermentation was scaled up to 10 L in a BioFlo 415 automatic fermentor.The dissolved oxygen was maintained at about 60%,and the recombinant cells were harvested at 7 h after induction of IPTG.Compared to the shaking-flask experiments,the ratio of target protein to total bacterium proteins increased about 1.5 times,and the enzyme activity increased from 46 U mL-1 to 78 U mL-1.These results laid foundation for the scale-up production of recombinant maleate cis-trans isomerase.Fig 7,Tab 2,Ref 17" @default.
- W2372233312 created "2016-06-24" @default.
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- W2372233312 date "2011-01-01" @default.
- W2372233312 modified "2023-09-25" @default.
- W2372233312 title "Optimization of Fermentation Conditions of Escherichia coli Strain Expressing a Recombinant Maleate cis-trans Isomerase" @default.
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