Influence of induction and operation mode on recombinant rhamnulose 1-phosphate aldolase production by Escherichia coli using the T5 promoter

Vidal, Luis; Ferrer, Pau; Álvaro, Gregorio; Benaiges, M. Dolors; Caminal, Glòria

doi:10.1016/j.jbiotec.2005.02.012

Cited by 34 publications

(34 citation statements)

References 31 publications

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“…10 and 11. The concentration and productivity of rh-GCSF show a colossal increase after the induction, which is in accordance with previous researches [11,13,20,22,23]. Increasing the initial glucose concentration from 10 to 20 g/l will increase the concentration of rh-GCSF from 950 to 1,750 mg/l; but when initial glucose concentration increases to 25 g/l, rh-GCSF concentration decreases significantly (1,570 mg/l).…”

Section: Optimization Of Glucose Concentration In the Selected Mediumsupporting

confidence: 91%

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Enhancement of human granulocyte-colony stimulating factor production in recombinant E. coli using batch cultivation

Babaeipour

Abbas

Sahebnazar

et al. 2009

Bioprocess Biosyst Eng

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Development of inexpensive and simple culture media is always favorable for recombinant protein over-expression in E. coli. The effects of medium composition on the production of recombinant human granulocyte-colony stimulating factor (rh-GCSF) were investigated in batch culture of E. coli BL21 (DE3) [pET23a-hgcsf]. First, the optimum medium for production of rh-GCSF was determined; and, then it was shown that mixture of amino acid addition at induction time, which was determined on the basis of amino acids frequency in the recombinant protein, increases recombinant protein expression level significantly. Furthermore, the effect of glucose concentration on productivity of rh-GCSF was investigated; 20 g/l of glucose will result in maximum attainable biomass and rh-GCSF in this process. At optimum conditions, a cell dry weight of 10.5 g/l, an expression level of about 35% of total cellular protein, rh-GCSF concentration of 1.75 +/- 0.1 g/l, and overall rh-GCSF yield of 165 +/- 5 mg/g were obtained.

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Section: Optimization Of Glucose Concentration In the Selected Mediumsupporting

confidence: 91%

“…Further increments of the initial glucose concentration of 20 g/l have an inhibitory effect on growth and cause a reduction in the specific growth rate. It has been reported that increasing the initial glucose concentration to 40-50 g/l will cause the cessation of growth [20][21][22][23].…”

Section: Optimization Of Glucose Concentration In the Selected Mediummentioning

confidence: 99%

Enhancement of human granulocyte-colony stimulating factor production in recombinant E. coli using batch cultivation

Babaeipour

Abbas

Sahebnazar

et al. 2009

Bioprocess Biosyst Eng

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“…The cultivation inoculum (10% v/v) was transferred to 250 mL Erlenmeyer flasks and kept under agitation at 200 rpm for 8 hours at 37° C. To understand the IPTG induction effect on growth and eIF antigen expression, cultures were induced by the addition of a final concentration of 1mM IPTG when optical density (OD 590nm ) reached 0.5 (early log phase) (20).…”

Section: Cultivation Conditionsmentioning

confidence: 99%

Influence of culture medium on the production of eif antigen from Leishmania chagasi in recombinant Escherichia coli

Vaz¹,

Franca²,

Andrade³

et al. 2011

Braz. J. Microbiol.

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With the advent of recombinant DNA technology, recombinant protein expression has become an important tool in the study of the structure, function and identification of new proteins, especially those with therapeutic functions. Escherichia coli has been the predominant prokaryote used in genetic engineering studies due to the abundance of information about its metabolism. Despite significant advances in molecular biology and immunology of infections, there are as yet no prophylactic drugs capable of preventing visceral leishmaniasis. It is therefore important to identify specific antigens in order to develop vaccines and diagnostic kits against this disease. The objective of this study was to evaluate the influence of culture medium on the production of eIF antigen from Leishmania chagasi in recombinant Escherichia coli. An induction procedure using IPTG was carried out in a series of trials, to observe the influence of culture medium (2xTY, TB) under expression of the recombinant eIF protein. Results showed that recombinant protein expression was associated to growth and that the highest eIF antigen expression was obtained in the 2xTY medium.

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“…After that, Vidal et al (15), switched to a commercially available expression system (Qiagen), consisting of the M15 strain and a two--plasmid system including the pQE40 expression vector, which uses the stronger T5 promoter. This system was used to overproduce a range of different aldolases, including a DHAP--dependent aldolase, the rhamnulose 1--phosphate aldolase (RhuA) (16).…”

Section: Introductionmentioning

confidence: 99%

Using promoter libraries to reduce metabolic burden due to plasmid-encoded proteins in recombinant Escherichia coli

et al. 2016

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Copyright and reuse:The Warwick Research Archive Portal (WRAP) makes this work by researchers of the University of Warwick available open access under the following conditions. Copyright © and all moral rights to the version of the paper presented here belong to the individual author(s) and/or other copyright owners. To the extent reasonable and practicable the material made available in WRAP has been checked for eligibility before being made available.Copies of full items can be used for personal research or study, educational, or not-for-profit purposes without prior permission or charge. Provided that the authors, title and full bibliographic details are credited, a hyperlink and/or URL is given for the original metadata page and the content is not changed in any way. A note on versions:The version presented here may differ from the published version or, version of record, if you wish to cite this item you are advised to consult the publisher's version. Please see the 'permanent WRAP URL' above for details on accessing the published version and note that access may require a subscription. AbstractThe over--expression of proteins in recombinant host cells often requires a significant amount of resources causing an increase in the metabolic load for the host. This results in a variety of physiological responses leading to altered growth parameters, including growth inhibition. Moreover, the expression of other plasmid--encoded genes such as antibiotic resistance genes or repressor proteins may also alter growth.In this work, we have developed a second generation of an Escherichia coli expression system with an antibiotic--free plasmid maintenance mechanism based on an auxotrophic marker (glyA). Metabolic burden related to plasmid maintenance and heterologous protein expression has been minimized by tuning the expression levels of a repressor protein (LacI) and glyA using a selected library of promoters and applying synthetic biology tools that allow the rapid construction of vectors. We apply our engineered antibiotic--free expression 2 system to the FucA over--production, showing increased production levels.Our results showed that the aforementioned approaches are of paramount importance in order to increment the protein production in terms of mass and activity. Acknowledgements:This work was supported by the project "Novel Alternatives for Microbial Production of Enzymes and Multienzymatic stereoselective Synthesis (EnzProSyn)". M. P. acknowledges the Universitat Autònoma de Barcelona for the pre--doctoral grant.

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Influence of induction and operation mode on recombinant rhamnulose 1-phosphate aldolase production by Escherichia coli using the T5 promoter

Cited by 34 publications

References 31 publications

Enhancement of human granulocyte-colony stimulating factor production in recombinant E. coli using batch cultivation

Enhancement of human granulocyte-colony stimulating factor production in recombinant E. coli using batch cultivation

Influence of culture medium on the production of eif antigen from Leishmania chagasi in recombinant Escherichia coli

Using promoter libraries to reduce metabolic burden due to plasmid-encoded proteins in recombinant Escherichia coli

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