Characterization and optimization of vascular endothelial growth factor165 (rhVEGF165) expression in Escherichia coli

Kang, Wonmo; Kim, S.; Lee, S.; Jeon, Eunyi; Lee, Y.; Yun, Ye-Rang; Suh, Chang Kook; Kim, H.W.; Jang, Jun‐Hyeog

doi:10.1016/j.pep.2012.10.004

Cited by 8 publications

(13 citation statements)

References 21 publications

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“…Recombinant human vascular endothelial growth factor (rhVEGF) was constructed using the pBAD‐HisA plasmid in Escherichia coli , as we previously described (Kang et al . ). Briefly, the open reading frame (ORF) encoding human VEGF was amplified from a cDNA library (Stratagene, La Jolla, CA).…”

Section: Methodsmentioning

confidence: 97%

Lysyl oxidase‐mediated VEGF‐induced differentiation and angiogenesis in human dental pulp cells

Bae

Park

et al. 2017

Int Endodontic J

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Section: Methodsmentioning

confidence: 97%

Lysyl oxidase‐mediated VEGF‐induced differentiation and angiogenesis in human dental pulp cells

Bae

Park

et al. 2017

Int Endodontic J

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“…However, only a small number of studies have shown the heterologous expression of VEGF in E . coli in the soluble protein fraction [ 38 , 54 ]. Our work is one of the rare cases in which VEGF is expressed in E .…”

Section: Discussionmentioning

confidence: 99%

“…The yield of recombinantly prepared VEGF mentioned in related studies varies on a case-by-case basis. Some studies do not present any yield [ 49 , 54 ] or the data are unclear [ 38 ]. In other works, the yield of purified active VEGF 121 or VEGF 165 is between 1 mg and 5 mg per L of bacterial culture [ 50 , 52 , 53 ], which is similar to the yield of mutant VEGF 121 purified in this work.…”

Section: Discussionmentioning

confidence: 99%

Enhanced Mitogenic Activity of Recombinant Human Vascular Endothelial Growth Factor VEGF121 Expressed in E. coli Origami B (DE3) with Molecular Chaperones

et al. 2016

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We describe the production of a highly-active mutant VEGF variant, α2-PI1-8-VEGF121, which contains a substrate sequence for factor XIIIa at the aminoterminus designed for incorporation into a fibrin gel. The α2-PI1-8-VEGF121 gene was synthesized, cloned into a pET-32a(+) vector and expressed in Escherichia coli Origami B (DE3) host cells. To increase the protein folding and the solubility, the resulting thioredoxin-α2-PI1-8-VEGF121 fusion protein was co-expressed with recombinant molecular chaperones GroES/EL encoded by independent plasmid pGro7.The fusion protein was purified from the soluble fraction of cytoplasmic proteins using affinity chromatography. After cleavage of the thioredoxin fusion part with thrombin, the target protein was purified by a second round of affinity chromatography. The yield of purified α2-PI1-8-VEGF121 was 1.4 mg per liter of the cell culture. The α2-PI1-8-VEGF121 expressed in this work increased the proliferation of endothelial cells 3.9–8.7 times in comparison with commercially-available recombinant VEGF121. This very high mitogenic activity may be caused by co-expression of the growth factor with molecular chaperones not previously used in VEGF production. At the same time, α2-PI1-8-VEGF121 did not elicit considerable inflammatory activation of human endothelial HUVEC cells and human monocyte-like THP-1 cells.

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“…[ 1 , 37 ] Kang et al. [ 3 ] optimized vascular endothelial growth factor 165 (rhVEGF 165 ) expression in E. coli and showed that shorter or longer induction time would decrease the expression level of rhVEGF 165 . The assumed reason probably was intracellular expression of the target protein which did not need to undergo a complicated translocation process.…”

Section: Resultsmentioning

confidence: 99%

“…[ 1,2 ] Induction conditions are very important for the expression of recombinant proteins. [ 3 ] Many studies on the optimization of induction conditions for the production of recombinant proteins have been reported. [ 4,5 ] Of all induction conditions, inducers play an important role in the expression of recombinant proteins.…”

Section: Introductionmentioning

confidence: 99%

Production of a new non-specific nuclease fromYersinia enterocoliticasubsp.palearctica: optimization of induction conditions using response surface methodology

Fang

Tang

et al. 2014

Biotechnology & Biotechnological Equipment

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A new non-specific nuclease from Yersinia enterocolitica subsp. palearctica (Y. NSN) was expressed in Escherichia coli (E. coli) BL 21 StarTM (DE3)plysS. Induction conditions, including isopropyl-β-D-thiogalactoside (IPTG) concentration, cell density (OD600), induction time and induction temperature, were optimized using response surface methodology. Statistical analysis of the results revealed that induction temperature and all the quadratic terms of variables had significant effects on enzyme activity of Y. NSN. The optimal induction conditions were as follows: 1.5 mmol/L IPTG, OD600 of 0.80, induction time of 20.5 h, and induction temperature of 32 °C. Under the optimized conditions, the highest enzyme activity could be obtained.

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Characterization and optimization of vascular endothelial growth factor165 (rhVEGF165) expression in Escherichia coli

Cited by 8 publications

References 21 publications

Lysyl oxidase‐mediated VEGF‐induced differentiation and angiogenesis in human dental pulp cells

Lysyl oxidase‐mediated VEGF‐induced differentiation and angiogenesis in human dental pulp cells

Enhanced Mitogenic Activity of Recombinant Human Vascular Endothelial Growth Factor VEGF121 Expressed in E. coli Origami B (DE3) with Molecular Chaperones

Production of a new non-specific nuclease fromYersinia enterocoliticasubsp.palearctica: optimization of induction conditions using response surface methodology

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