Microbial Expression Systems and Manufacturing from a Market and Economic Perspective

Meyer, Hans‐Peter; Schmidhalter, Diego R.

doi:10.5772/29417

Cited by 13 publications

(9 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A large number of molecular cloning techniques and genetic tools have been developed using E. coli. Rapid strain development allowed by these techniques can significantly reduce costs for industrial development (Meyer and Schmidhalter, 2012).…”

Section: Why E Coli?mentioning

confidence: 99%

“…coli has been used as the host for industrial production of various chemicals, including trypotophan, phenylalanine, threonine, lysine, nucleotides, succinate, itaconic acid, polyhydroxybutyate, and 1,3-PDO, as discussed below. In terms of biopharmaceutical applications, it was reported that of more than 100 recombinant protein products in the US and European markets, 34% of these are expressed in E. coli (Meyer and Schmidhalter, 2012). In 1998, E. coli K strain MG-1655 was sequenced (Blattner et al, 1997).…”

Section: Why E Coli?mentioning

confidence: 99%

“…However, BL21 (B strain) and its derivative BL21(DE3), containing λDE3 lysogen harboring the gene for T7 RNA polymerase, are popular for recombinant protein production (Studier and Moffatt, 1986). Applications that involve high protein expression, such as production of protein biopharmaceuticals, most frequently use BL21 as a host because of its lack of lon and ompT proteases, as well as its insensitivity to high glucose concentrations (Meyer and Schmidhalter, 2012). The insensitivity to glucose concentration stems from high activity of the glyoxylate shunt, gluconeogenesis, anaplerotic pathways, and the TCA cycle, leading to reduced acetate production and efficient glucose utilization (Phue et al, 2008).…”

Section: Why E Coli?mentioning

confidence: 99%

See 2 more Smart Citations

Escherichia coli as a host for metabolic engineering

Pontrelli

Chiu

Lan

et al. 2018

Metabolic Engineering

290

140

View full text Add to dashboard Cite

Over the past century, Escherichia coli has become one of the best studied organisms on earth. Features such as genetic tractability, favorable growth conditions, well characterized biochemistry and physiology, and availability of versatile genetic manipulation tools make E. coli an ideal platform host for development of industrially viable productions. In this review, we discuss the physiological attributes of E. coli that are most relevant for metabolic engineering, as well as emerging techniques that enable efficient phenotype construction. Further, we summarize the large number of native and non-native products that have been synthesized by E. coli, and address some of the future challenges in broadening substrate range and fighting phage infection.

show abstract

Section: Why E Coli?mentioning

confidence: 99%

Section: Why E Coli?mentioning

confidence: 99%

Section: Why E Coli?mentioning

confidence: 99%

See 1 more Smart Citation

Escherichia coli as a host for metabolic engineering

Pontrelli

Chiu

Lan

et al. 2018

Metabolic Engineering

290

140

View full text Add to dashboard Cite

show abstract

“…Nearly 30% of currently approved recombinant therapeutic proteins are produced in E. coli (Huang et al 2012) as the host of choice due to its rapid growth on simple inexpensive media, high-yield production and well-known physiology and genetics (Swartz, 2001;Meyer and Schmidhalter, 2012). Proteins of therapeutic value, including antibodies and antibody fragments, often contain disulfide bond bridges that are necessary for their folding, stability and/or function.…”

Section: Introductionmentioning

confidence: 99%

“…coli was the first host used to produce recombinant DNA (rDNA) pharmaceuticals, enabling the approval of recombinant DNA (rDNA) human insulin (Humulin ® , licensed by GENENTECH to ELI LILLY) in 1982 and marketing of Monsanto's bovine growth hormone (bGH) in 1994 (Swartz, 2001;Meyer and Schmidhalter, 2012).…”

Section: Introductionmentioning

confidence: 99%

Production of antibody fragment (Fab) throughout Escherichia coli fed-batch fermentation process: Changes in titre, location and form of product

Jalalirad¹

2013

Electron. J. Biotechnol.

View full text Add to dashboard Cite

Background: Recombinant proteins, including antibodies and antibody fragments, often contain disulfide bond bridges that are necessary for their folding, stability and function. Production of disulfidebond-containing proteins in the periplasm of Escherichia coli has been very useful, due to unique characteristics of the periplasm, for obtaining fully active and correctly folded products and for alleviating downstream processing. Results: In this study, fed-batch cultivation of Escherichia coli (E. coli) for production of Fab D1.3, which is an anti-hen egg white lysozyme (HEWL) antibody fragment was carried out at 37ºC, and the bacterial cells were induced by adding 0.1 mM IPTG to the culture medium. Fermentor was sampled over the course of fermentation; the bacterial cells were centrifugally separated from the culture broth and subjected to osmotic shock (with excluding HEWL) and sonication procedures. The resulting fractions were analysed for Fab using a combination of ELISA, SDS-PAGE and Western blotting and changes in product titre, location, and form was assessed throughout growth. It was shown that osmotic shock released the Fab from the periplasm very efficiently and its efficacy was 20-45% more than sonication. This study demonstrates that, at high cell density cultivation in fermentor, target product can appear inside and outside the cells, depending on the time of induction. The maximum amount of Fab (47 mg/l) in the periplasm was reached at 14 hrs cultivation (4 hrs post induction), being suitable time for cell harvest, selective periplasmic extraction and downstream capture. The Fab increasingly leaked into the culture medium, and reached its maximum culture medium titre of ~78 mg/l after 6 hrs post induction. After 16 hrs cultivation (6 hrs post induction) the amount of Fab remained constant in different locations within and outside the cells. Western blot analysis of cell fractions showed that certain amount of the Fab was also produced in the cells as insoluble form. Conclusions: In this work we showed that the production of Fab in the periplasm during high cell density cultivation of E. coli in fermentor can be challenging as the product may appear in various locations within and outside the cells. To exploit the advantages of the periplasmic expression systems for purification in downstream processing, bacterial cells should be harvested when they maintain.

show abstract

The History and Economic Relevance of Industrial Scale Suspension Culture of Living Cells

Meyer¹,

Schmidhalter²

2014

Industrial Scale Suspension Culture of Living Cells

View full text Add to dashboard Cite

Microbial Expression Systems and Manufacturing from a Market and Economic Perspective

Cited by 13 publications

References 44 publications

Escherichia coli as a host for metabolic engineering

Escherichia coli as a host for metabolic engineering

Production of antibody fragment (Fab) throughout Escherichia coli fed-batch fermentation process: Changes in titre, location and form of product

The History and Economic Relevance of Industrial Scale Suspension Culture of Living Cells

Contact Info

Product

Resources

About