Cost analysis based on bioreactor cultivation conditions: Production of a soluble recombinant protein using Escherichia coli BL21(DE3)

Cardoso, Valdemir M.; Campani, Gilson; Santos, M. P.; Silva, Gabriel G.; Pires, Manuella C.; Gonçalves, Viviane Maimoni; Giordano, Roberto de Campos; Sargo, Cíntia Regina; Horta, Antonio Carlos Luperni; Zangirolami, Teresa Cristina

doi:10.1016/j.btre.2020.e00441

Cited by 59 publications

(46 citation statements)

References 90 publications

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“…The rising demand for biotechnological products as well as for the development of new bioprocesses requires continuous studies on microbial physiology and population growth [1,2]. In recent years, numerous studies on the optimization of cultivation conditions (e.g., medium composition, temperature, pH level) were conducted, which improved the mathematical description of bioprocesses [3][4][5][6][7].…”

Section: Introductionmentioning

confidence: 99%

Single Mathematical Parameter for Evaluation of the Microorganisms’ Growth as the Objective Function in the Optimization by the DOE Techniques

et al. 2020

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The cultivation of bacteria sets a ground for studying biological processes in many scientific disciplines. The development of the bacterial population is commonly described with three factors that can be used to evaluate culture conditions. However, selecting only one of them for the optimization protocol is rather problematic and may lead to unintended errors. Therefore, we proposed a novel mathematical approach to obtain a single factor that could be used as the objective function to evaluate the whole growth dynamic and support the optimization of the biomass production process. The sigmoidal-shape curve, which is the commonly used function to plot the amount of biomass versus time, was the base for the mathematical analysis. The key process parameters, such as maximal specific growth rate and lag-phase duration were established with the use of mathematical coefficients of the model curve and combined to create the single growth parameter. Moreover, this parameter was used for the exemplary optimization of the cultivation conditions of Klebsiella pneumoniae that was cultured to be further used in the production of lytic bacteriophages. The proposed growth parameter was successfully validated and used to calculate the optimal process temperature of the selected bacterial strain. The obtained results indicated that the proposed mathematical approach could be effortlessly adapted for a precise evaluation of growth curves.

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Section: Introductionmentioning

confidence: 99%

Single Mathematical Parameter for Evaluation of the Microorganisms’ Growth as the Objective Function in the Optimization by the DOE Techniques

et al. 2020

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confidence: 99%

“…Complex media additives are usually also cost-intensive and media cost was shown to be one of the core expenses in bioreactor cultivations [25,26]. Media cost was mainly in uenced by the feedstock price for peptones [25], which suggests that the omittance of complex media additives is bene cial for industrial applications. Additionally, Yang et al (2020) reported that these components can disguise the true product yield, since isopropanol production reached 112 % of the maximal theoretical yield [17].…”

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confidence: 99%

Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

Novak

Kutscha

Pflügl

2020

Preprint

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Background: Acetate is an abundant carbon source and its use as an alternative feedstock has great potential for the production of fuel and platform chemicals. Acetoin and 2,3-butanediol represent two of these potential platform chemicals.Results: The aim of this study was to produce 2,3-butanediol and acetoin from acetate in Escherichia coli W. The key strategies to achieve this goal were: strain engineering, in detail the deletion of mixed acid fermentation pathways E. coli W ΔldhA ΔadhE Δpta ΔfrdA 445_Ediss and the development of a new defined medium containing five amino acids and seven vitamins. Stepwise reduction of the media additives further revealed that diol production from acetate is mediated by the availability of aspartate. Other amino acids or TCA cycle intermediates did not enable growth on acetate. Cultivation under controlled conditions in batch and pulsed fed-batch experiments showed that aspartate was consumed before acetate, indicating that co-utilization is not a prerequisite for diol production. The addition of aspartate gave cultures a start-kick and was not required for feeding. Pulsed fed-batches resulted in the production of 1.43 g l-1 from aspartate and acetate and 1.16 g l-1 diols (2,3-butanediol and acetoin) from acetate alone. The yield reached 0.09 g diols per g acetate, which accounts for 26 % of the theoretical maximum.Conclusion: This study for the first time showed acetoin and 2,3-butanediol production from acetate as well as the use of chemically defined medium for product formation from acetate in E. coli. Hereby, we provide a solid base for process intensification and the investigation of other potential products.

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mentioning

confidence: 99%

“…Complex media additives are usually also cost-intensive and media cost was shown to be one of the core expenses in bioreactor cultivations (24,25). Media cost was mainly in uenced by the feedstock price for peptones (24), which suggests that the omittance of complex media additives is bene cial for industrial applications. Additionally, Yang et al (2020) reported that these components can disguise the true product yield, since isopropanol production reached 112 % of the maximal theoretical yield (16).…”

mentioning

confidence: 99%

Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

Novak

Kutscha

Pflügl

2020

Preprint

View full text Add to dashboard Cite

Background Acetate is an abundant carbon source and its use as an alternative feedstock has great potential for the production of fuel and platform chemicals. Acetoin and 2,3-butanediol represent two of these potential platform chemicals. Results The aim of this study was to produce 2,3-butanediol and acetoin from acetate in Escherichia coli W. The key strategies to achieve this goal were: strain engineering, in detail the deletion of mixed acid fermentation pathways E. coli W ΔldhA ΔadhE Δpta ΔfrdA 445_Ediss and the development of a new defined medium containing five amino acids and seven vitamins. Stepwise reduction of the media additives further revealed that diol production from acetate is mediated by the availability of aspartate. Other amino acids or TCA cycle intermediates did not enable growth on acetate. Cultivation under controlled conditions in batch and pulsed fed-batch experiments showed that aspartate was consumed before acetate, indicating that co-utilization is not a prerequisite for diol production. The addition of aspartate gave cultures a start-kick and was not required for feeding. Pulsed fed-batches resulted in the production of 1.43 g l-1 from aspartate and acetate and 1.16 g l-1 diols (2,3-butanediol and acetoin) from acetate alone. The yield reached 0.09 g diols per g acetate, which accounts for 26 % of the theoretical maximum.Conclusion This study for the first time showed acetoin and 2,3-butanediol production from acetate as well as the use of chemically defined medium for product formation from acetate in E. coli. Hereby, we provide a solid base for process intensification and the investigation of other potential products.

show abstract

Cost analysis based on bioreactor cultivation conditions: Production of a soluble recombinant protein using Escherichia coli BL21(DE3)

Cited by 59 publications

References 90 publications

Single Mathematical Parameter for Evaluation of the Microorganisms’ Growth as the Objective Function in the Optimization by the DOE Techniques

Single Mathematical Parameter for Evaluation of the Microorganisms’ Growth as the Objective Function in the Optimization by the DOE Techniques

Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

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