High Yield Production of Heterologous Proteins with Escherichia coli

Tripathi, Nagesh K.

doi:10.14429/dsj.59.1501

Cited by 69 publications

(52 citation statements)

References 74 publications

(133 reference statements)

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“…Defined media are generally used to attain high cell density, as the nutrient concentrations are well-known and can be controlled during culture. However, complex or semi-defined media are sometimes necessary to boost product formation [35]. We had selected four complex media i.e Luria Broth, Nutrient Broth, YT media and SOB, with minimal media as control to study their effect on lipase production from our recombinant culture.…”

Section: Firstmentioning

confidence: 99%

Studies on Recombinant Lipase Production by E. Coli: Effect of Media And Bacterial Expression System Optimization

Kaur¹

2017

IJMBOA

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Recombinant protein production is a vast field growing larger in each passing year due to continuous emergence of new host, vectors and techniques for cloning and expression. Earlier, a Bacillus lipase gene was cloned in pGEMT/ E. coli DH5α with 12 units/ml lipase production. In the present investigation, attempt has been made to optimize the production of recombinant lipase by E. coli by varying different parameters. Amongst the different strains of E. coli, M46 cells were observed to be the best host with 19 units/ml lipase production. Addition of surfactant and changing media to nutrient broth lead to 2.8 fold and 1.3 fold respective increase in lipase production. Amongst different vectors, pET 28a vector showed the highest total lipase production but it proved toxic for the cells, while pQE30-UA vector gives 5 fold enhanced total lipase production with more stable expression. With the combination of optimized host, vector, surfactant and media, 18 fold increases in lipase production (214 units/ml) was achieved.

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Section: Firstmentioning

confidence: 99%

Studies on Recombinant Lipase Production by E. Coli: Effect of Media And Bacterial Expression System Optimization

Kaur¹

2017

IJMBOA

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“…Group 1 also includes 85 % of the experimental points from the cultivations carried out with E. coli "a" and "d" in complex media. It has been suggested in the literature that medium supplementation with amino acids or proteins can partially relieve the stress caused by the recombinant protein production (Shiloach and Fass, 2005;Shojaosadati et al, 2008;Tripathi et al, 2009). On the other hand, Group 2 consists mainly of data from E. coli "b" and "c" cultures, which were carried out in defined media.…”

Section: Correlation Between Permittivity and Cell Concentration -Cx mentioning

confidence: 99%

On-Line Monitoring of Biomass Concentration Based on a Capacitance Sensor: Assessing the Methodology for Different Bacteria and Yeast High Cell Density Fed-Batch Cultures

Horta

Silva

Sargo

et al. 2015

Braz. J. Chem. Eng.

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-The performance of an in-situ capacitance sensor for on-line monitoring of biomass concentration was evaluated for some of the most important microorganisms in the biotechnology industry: Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris and Bacillus megaterium. A total of 33 batch and fed-batch cultures were carried out in a bench-scale bioreactor and biomass formation trends were followed by dielectric measurements during the growth phase as well as the induction phase, for 5 recombinant E. coli strains. Permittivity measurements and viable cellular concentrations presented a linear correlation for all the studied conditions. In addition, the permittivity signal was further used for inference of the cellular growth rate. The estimated specific growth rates mirrored the main trends of the metabolic states of the different cells and they can be further used for setting-up control strategies in fed-batch cultures.

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“…The pH was controlled (on/off) at 6.7 (pHmeter GLI PRO) by addition of NH 4 OH (30%) in the batch phase and at 6.9 in the feeding phase. The upward shift in the pH was to ensure an adequate nitrogen supply for the growing cell population and avoid the possibility that growth and heterologous protein production would become limited by nitrogen shortage (Seeger et al, 1995;Korz et al, 1995;Lee, 1996;Babaeipour et al, 2007;Carvalho et al, 2012;Tripathi et al, 2009). Temperature was set at 25, 30, 35 or 37 °C (depending on the strain being cultivated).…”

Section: Bioreactor Operation and Instrumentationmentioning

confidence: 99%

“…coli) are used in the production of numerous products of the pharmaceutical industry like insulin, growth hormones, vaccines and interferons (Liljeqvist and Stahl, 1999;Eiteman and Altman, 2006;Tripathi et al, 2009). Regardless of the product of interest, the cultures are carried out with the initial goal of achieving high cell density and then proceed to the induction of the synthesis of the desired protein.…”

Section: Introductionmentioning

confidence: 99%

A supervision and control tool based on artificial intelligence for high cell density cultivations

Horta

Silva

Sargo

et al. 2014

Braz. J. Chem. Eng.

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-High cell density cultivations of recombinant E. coli have been increasingly used for the production of heterologous proteins. However, it is a challenge to maintain these cultivations within the desired conditions, given that some variables such as dissolved oxygen concentration (DOC) and feed flow rate are difficult to control. This paper describes the software SUPERSYS_HCDC, a tool developed to supervise fed-batch cultures of rE. coli with biomass concentrations up to 150 g DCW /L and cell productivities up to 9 g DCW .L -1 .h -1 . The tool includes automatic control of the DOC by integrated action of the stirrer speed as well as of the air and oxygen flow rates; automatic start-up of the feed flow of fresh medium (system based on a neural network committee); and automatic slowdown of feeding when oxygen consumption exceeds the maximum capacity of the oxygen supply.

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High Yield Production of Heterologous Proteins with Escherichia coli

Cited by 69 publications

References 74 publications

Studies on Recombinant Lipase Production by E. Coli: Effect of Media And Bacterial Expression System Optimization

Studies on Recombinant Lipase Production by E. Coli: Effect of Media And Bacterial Expression System Optimization

On-Line Monitoring of Biomass Concentration Based on a Capacitance Sensor: Assessing the Methodology for Different Bacteria and Yeast High Cell Density Fed-Batch Cultures

A supervision and control tool based on artificial intelligence for high cell density cultivations

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