Cloning and Expression of Human Stem Cell Factor Fused with Thrombopoietin Mimetic Peptide in Escherichia coli

Su, Lin; Chen, Songsen; Yang, Kailun; Liu, Changzheng; Liang, Zhaoming

doi:10.1007/s10529-006-9017-7

Cited by 4 publications

(7 citation statements)

References 13 publications

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“…Inclusion body formation, as might be seen in the insoluble fraction of the cell lysate, is hardly observed. Up to the present, SCF expression in E. coli has always led to insoluble inclusion bodies (Langley et al, 1992;Lili et al, 2006;Potala and Verma, 2010;Su et al, 2006;Wang et al, 2008), thus the formation of soluble TRX-mSCF is obviously due to the TRX presence.…”

Section: Expression Of Trx-mscfmentioning

confidence: 86%

“…The simultaneous renaturation and purification of E. coli-derived human SCF has been described as well (Lili et al, 2006;Wang et al, 2008). Furthermore, several fusion constructs of human and murine SCF have been reported (diphtheria toxin-SCF (Potala and Verma, 2010), TPO-SCF (Su et al, 2006), TPO-SCF-thioredoxin fusionprotein (Zang et al, 2007) and TRX-SCF (LaVallie et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

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Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Bals

Schambach

Meyer

et al. 2011

Journal of Biotechnology

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Section: Expression Of Trx-mscfmentioning

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Bals

Schambach

Meyer

et al. 2011

Journal of Biotechnology

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“…It also includes cIts875, a gene encoding a temperature-sensitive regulation protein, upstream of the promoter, and a strong downstream transcription stop sequence. Although pBV220 has been used to successfully express some medicinal proteins and enzymes on a laboratory scale (21,34,38,41), this is its first documented use in an industrial process.…”

Section: Discussionmentioning

confidence: 99%

Microbial Conversion of Glycerol to 1,3-Propanediol by an Engineered Strain of Escherichia coli

Tang

Tan

Hong

et al. 2009

Appl Environ Microbiol

143

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In an effort to improve industrial production of 1,3-propanediol (1,3-PD), we engineered a novel polycistronic operon under the control of the temperature-sensitive lambda phage P L P R promoter regulated by the cIts857 repressor and expressed it in Escherichia coli K-12 ER2925. The genes for the production of 1,3-PD in Clostridium butyricum, dhaB1 and dhaB2, which encode the vitamin B 12 -independent glycerol dehydratase DhaB1 and its activating factor, DhaB2, respectively, were tandemly arrayed with the E. coli yqhD gene, which encodes the 1,3-propanediol oxidoreductase isoenzyme YqhD, an NADP-dependent dehydrogenase that can directly convert glycerol to 1,3-PD. The microbial conversion of 1,3-PD from glycerol by this recombinant E. coli strain was studied in a two-stage fermentation process. During the first stage, a novel high-cell-density fermentation step, there was significant cell growth and the majority of the metabolites produced were organic acids, mainly acetate. During the second stage, glycerol from the fresh medium was rapidly converted to 1,3-PD following a temperature shift from 30°C to 42°C. The by-products were mainly pyruvate and acetate. During this two-stage process, the overall 1,3-PD yield and productivity reached 104.4 g/liter and 2.61 g/liter/h, respectively, and the conversion rate of glycerol to 1,3-PD reached 90.2% (g/g). To our knowledge, this is the highest reported yield and productivity efficiency of 1,3-PD with glycerol as the sole source of carbon. Furthermore, the overall fermentation time was only 40 h, shorter than that of any other reports.

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“…The expression level in the batch was 207 mg/L of culture supernatant whose level was improved by 1.7‐fold, as compared that of laboratory‐scale experiments . The rhSCF was expressed as IBs in E. coli .…”

Section: Discussionmentioning

confidence: 99%

“…rhSCF has been expressed in E. coli as soluble and insoluble forms on a laboratory and pilot scale by many laboratories . The active rhSCF from IBs involves cell lysis, extraction and cleaning of IBs, solubilization of IBs, refolding into its native conformation, and purification procedure . Recently, Akuta et al.…”

Section: Discussionmentioning

confidence: 99%

Large‐scale purification and characterization of recombinant human stem cell factor in Escherichia coli

Chen

Cai

Zhang

et al. 2017

Biotech and App Biochem

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The pharmacological importance of recombinant human stem cell factor (rhSCF) has increased the demand to establish effective and large-scale production and purification processes. A good source of bioactive recombinant protein with capability of being scaled-up without losing activity has always been a challenge. The objectives of the study were the rapid and efficient pilot-scale expression and purification of rhSCF. The gene encoding stem cell factor (SCF) was cloned into pBV220 and transformed into Escherichia coli. The recombinant SCF was expressed and isolated using a procedure consisting of isolation of inclusion bodies (IBs), denaturation, and refolding followed by chromatographic steps toward purification. The yield of rhSCF reached 835.6 g/20 L, and the expression levels of rhSCF were about 33.9% of the total E. coli protein content. rhSCF was purified by isolation of IBs, denaturation, and refolding, followed by SP-Sepharose chromatography, Source 30 reversed-phase chromatography, and Q-Sepharose chromatography. This procedure was developed to isolate 5.5 g of rhSCF (99.5% purity) with specific activity at 0.96 × 10 IU/mg, endotoxin levels of pyrogen at 1.0 EU/mg, and bacterial DNA at 10 ng/mg. Pilot-scale fermentations and purifications were set up for the production of rhSCF that can be upscaled for industry.

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Cloning and Expression of Human Stem Cell Factor Fused with Thrombopoietin Mimetic Peptide in Escherichia coli

Cited by 4 publications

References 13 publications

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Microbial Conversion of Glycerol to 1,3-Propanediol by an Engineered Strain of Escherichia coli

Large‐scale purification and characterization of recombinant human stem cell factor in Escherichia coli

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