Optimization of fusion proinsulin production by high cell-density fermentation of recombinantE. coli

Bae, Cheon-Soon; Hong, Minsun; Chang, Seung-Gu; Kim, Dae-Young; Shin, Hang-Cheol

doi:10.1007/bf02932459

Cited by 12 publications

(6 citation statements)

References 17 publications

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“…So, importantly, the addition of YE in an exceeded concentration of over 10 g/L could minimize acetate production pathway [7]. When YE concentration was increased from 0 to 10 g/L, it effected to the increase in specific growth rate (μ) of E. coli, but the level of YE over 10 g/L did not [31]. This result was confirmed by [32] who reported the decrease in YE/glucose fraction did not affect growth kinetics, but reduced the final cell concentration by about 10%, and also reduced the cell yield coefficient.…”

Section: The Addition Of Yeast Extract As a Supplement To Enhance The...mentioning

confidence: 99%

High Cell Density Cultivation (HCDC) of E. coli with Intensive Multiple Sequential Batches Together with Fed-Batch at Cell Level (FBC) Superior to the Fed-Batch Technique

Chamsart

2022

Preprint

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High cell density cultivation (HCDC) is a technique that was developed and used in the cultivation of various microorganisms. Escherichia coli is the most prominent in the bacterial group and popular at the industrial scale for the production of bio-products. The HCDC conducted with the Fed-Batch modes is complicated. In this study, we developed a batch production medium (BPM), a minimal defined medium for industrial utilization with the potential use of dextrin at high concentrations as the major carbon source. Dextrin prevents cell cultivation system from the Crabtree effect due to its gradual hydrolysis to release glucose and gradual assimilation into E. coli cells like Fed-Batch at Cell Level (FBC). Yeast extract was added into the BPM to enhance cell growth. It was operated using 5-L bioreactors with the aerobic batch cultivation at high concentrations of dextrin, i.e., 90 g/L to achieve high-cell-density concentration at a high production rate superior to the Fed-Batch technique. Further, the Intensive Multiple Sequential Batch (IMFB) technique was developed to achieve a high yield at a high productivity. The maximum values of cell biomass, x; specific growth rate\(, \mu\); yield coefficient, Yx/s; productivity, rx; and efficiency (Ef) were 46.31 g/L, 0.54 h− 1, 0.55 g/g, 3.86 g/L/h and 98%, respectively. The results showed that cell cultivations and growth kinetics, biochemical engineering mass balances, and design of impeller speeds using fluid dynamics in the 5-L bioreactor of aerobic batch cell cultivation for the production of E. coli cell biomass a high-density concentration using dextrin to control the FBC at a high titer can be used for the scale-up of industrial bioreactors.

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Section: The Addition Of Yeast Extract As a Supplement To Enhance The...mentioning

confidence: 99%

High Cell Density Cultivation (HCDC) of E. coli with Intensive Multiple Sequential Batches Together with Fed-Batch at Cell Level (FBC) Superior to the Fed-Batch Technique

Chamsart

2022

Preprint

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“…addition of 1% L-arabinose for all cases, which could have resulted from the utilization of cellular machinery for the expression of target protein, salmosin, instead of the cell growth [1]. The induction at the early stage during the fedbatch fermentation significantly inhibited the cell growth of recombinant E. coli, with DCW of 45.18 g/l.…”

Section: High Cell Density Fermentation For Salmosin Production 1055mentioning

confidence: 99%

“…Our results indicate that the cell growth of recombinant E. coli is strongly influenced by the induction time. In addition, the induction was proved to be preferably done in the late stage of the cell growth phase for the achievement of high cell density of recombinant E. coli [1].…”

Section: High Cell Density Fermentation For Salmosin Production 1055mentioning

confidence: 99%

“…We focused on the maximization of cell density of recombinant E. coli, based on the hypothesis that the production yield of expressed inclusion body is dependent on the cell density, which is therefore one of the major requirements for the high productivity of recombinant protein [2,24]. To determine the optimal nitrogen sources for the high cell density, the cultures were prepared in a 500 ml flask with various ratios of yeast extract to bactotryptone, which is typically used in the E. coli culture, since one or more nitrogen sources usually increases the cell density as well as the cell growth rate [1]. As the results, the maximum cell density was observed with the ratio of 3% yeast extract to 1% bacto-tryptone (Supplementary Table S1).…”

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confidence: 99%

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Production of a Platelet Aggregation Inhibitor, Salmosin, by High Cell Density Fermentation of Recombinant Escherichia coli

Seo

Choi²,

Chung³

et al. 2011

J. Microbiol. Biotechnol.

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Optimal conditions for a high cell density fermentation were investigated in a recombinant Escherichia coli producing salmosin, a platelet aggregation inhibitor. The optimized carbon and nitrogen sources were glycerol 10 g/l, yeast extract 30 g/l, and bacto-tryptone 10 g/l, yielding the dry cell weight (DCW) of 10.61 g/l in a 500 ml flask culture. The late-stage induction with 1% L-arabinose in a 5 l jar fermentor showed the highest DCW of 65.70 g/l after 27 h of the fed-batch fermentation. Around 2,200 mg/l of the protein was expressed as an inclusion body that was then refolded to obtain the active salmosin of 96 mg/l. We also confirmed the inhibitory activity against platelet aggregation of the active salmosin from the high cell density fermentation.

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“…Specifically considering cloning, recombinant protein production 15 is a typical example where genetically modified cells must be isolated individually and expanded in culture. Each of the single-cell clones is then screened to select for the best protein producer candidate.…”

Section: Introductionmentioning

confidence: 99%

Impedance-Based Single-Cell Pipetting

et al. 2020

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Optimization of fusion proinsulin production by high cell-density fermentation of recombinantE. coli

Cited by 12 publications

References 17 publications

High Cell Density Cultivation (HCDC) of E. coli with Intensive Multiple Sequential Batches Together with Fed-Batch at Cell Level (FBC) Superior to the Fed-Batch Technique

High Cell Density Cultivation (HCDC) of E. coli with Intensive Multiple Sequential Batches Together with Fed-Batch at Cell Level (FBC) Superior to the Fed-Batch Technique

Production of a Platelet Aggregation Inhibitor, Salmosin, by High Cell Density Fermentation of Recombinant Escherichia coli

Impedance-Based Single-Cell Pipetting

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