Effect of IPTG Concentration on Recombinant Human Prethrombin-2 Expression in<i>Escherichia coli</i>BL21(DE3) ArcticExpress

Silaban, Saronom; Gaffar, Shabarni; Simorangkir, Murniaty; Maksum, Iman Permana; Subroto, Toto

doi:10.1088/1755-1315/217/1/012039

Cited by 18 publications

(15 citation statements)

References 19 publications

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“…Poultry needs large amounts of vitamins since they cannot synthesize vitamins for the ongoing reactions of metabolism in the body. [21][22][23] Normal albumin level in chicken eggs are about 4.0 to 5.9 g / dL. It is concluded that the albumin levels after pyridoxine supplementation by each method is increased compared to the normal levels.…”

Section: Analysis Of Chicken Egg Globulin Protein Levelsmentioning

confidence: 95%

Effective Methods of Pyridoxine Supplementation in Laying Hens to Albumin and Globulin Levels

Silaban¹,

Sinaga²,

Damanik³

et al. 2020

RJC

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This research investigates the effective methods of pyridoxine supplementation to enhance the protein (albumin and globulin) level of chicken egg. 12 laying hens that been ready to produce eggs were categorized into three groups based on pyridoxine supplementation methods (via drinking water, ration, and intravenous injection). Each group received supplementation pyridoxine treatment with dosage of 3 mg/kg ransoms for 68 days. The albumin and globulin levels were analyzed using Folin's Fenol technique and compared to the standard egg. We found that only albumin level could be enhanced. The intravenous injection was the most effective method to significantly increase the albumin levels compared.

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Section: Analysis Of Chicken Egg Globulin Protein Levelsmentioning

confidence: 95%

Effective Methods of Pyridoxine Supplementation in Laying Hens to Albumin and Globulin Levels

Silaban¹,

Sinaga²,

Damanik³

et al. 2020

RJC

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“…IPTG is the inducer for gene expression from the T7 promoter. It regulates the gene expression at the transcription level and affects the accumulation level of biomass (Silaban, Gaffar, Simorangkir, Maksum, & Subroto, 2019). Excessively high concentrations of IPTG have potential toxicity (Rizkia et al, 2015).…”

Section: Effect Of the Protein Expression Level On The Pb 2+ Adsorpmentioning

confidence: 99%

Display of lead‐binding proteins on Escherichia coli surface for lead bioremediation

Jia

Liu

et al. 2020

Biotech & Bioengineering

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Cell surface display of heavy metal-binding proteins has been used to enhance the adsorption capacity of heavy metals and the engineered microbial cells can be potentially used for the bioremediation of heavy metals. In this study, the proteins PbrR, PbrR691, and PbrD from the Cupriavidus metallidurans strain CH34 were displayed on the extracellular membrane of Escherichia coli BL21 cells, with the N-domain of ice-nucleation protein as the anchor protein to achieve specific adsorption of lead ions (Pb 2+) and bioremediation of lead in the soil. The localization of fusion proteins was confirmed by western blot analysis. We investigated the effects of fusion pattern, expression level, heavy metal concentration, and the presence of other heavy metal ions on the adsorption of Pb 2+ by these engineered bacteria, and the optimal linker peptide (flexible linker) and inducer concentration (0.5 mM) were obtained. The engineered bacteria showed specific selectivity and strong adsorption capacity for Pb 2+. The maximum Pb 2+ adsorption capacity of strains displaying the three proteins (PbrR, PbrR691, and PbrD) were 942.1-, 754.3-, and 864.8-μmol/g cell dry weight, respectively, which was the highest reported to date. The engineered E. coli bacteria were also applied to Pb 2+-contaminated soil and the detoxification effects were observed via the seed germination test and the growth of Nicotiana benthamiana in comparison with the control BL21, which provides the proof-ofconcept for in situ remediations of Pb 2+-contaminated water or soil.

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“…Inclusion bodies are protein aggregate that consists of partially folded and misfolded protein that interacts via hydrophobic interaction. Their formation is often undesirable because they generally lack biological activity (Gomes et al, 2016;Silaban et al, 2019;Singh and Panda, 2005). This approach can give the advantage by refolding the inactive protein in inclusion bodies into its active form (Upadhyay et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Expression of human epidermal growth factor using Ssp DnaB mini-intein as fusion partner in Escherichia coli BL21(DE3)

Yosua¹,

Rima²,

Sriwidodo³

et al. 2021

J Appl Pharm Sci

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The human epidermal growth factor (hEGF) is widely used clinically as a wound healer, as it has a vital role in stimulating cell proliferation, differentiation, and migration. Consequently, the large-scale production of recombinant hEGF in E. coli has been developed to meet the high demand for hEGF clinically. However, intracellular proteins in E. coli, especially small proteins like hEGF, are degraded by proteases. To overcome this issue, hEGF was fused with CBD-Ssp DnaB to construct a fusion protein CBD-Ssp DnaB-hEGF. This study was conducted to obtain refolded hEGF from the inclusion bodies of CBD-Ssp DnaB-hEGF. The experiment was carried out using E. coli BL21(DE3) containing plasmid pD861-CBD-Ssp DnaB-hEGF. The CBD-Ssp DnaB-hEGF gene was constructed by fusing CBD-Ssp DnaB and hEGF gene and then was optimized. The method was started with E. coli transformation, CBD-Ssp DnaB-hEGF expression, inclusion bodies solubilization, refolding, and simultaneous cleavage to release hEGF. The CBD-Ssp DnaB-hEGF was expressed as inclusion bodies, which can then be purified by washing with Triton X-100 and 1 M urea. The inclusion bodies were solubilized in 8 M urea, the solubilized CBD-Ssp DnaB-hEGF was reformed by dialysis, and then hEGF was spliced by shifting the pH from 8.5 to 6.0 to yield a concentration of 0.163 mg/ml. Therefore, we concluded that hEGF was obtained from the solubilized CBD-Ssp DnaB-hEGF from inclusion bodies produced by E. coli BL21(DE3).

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Effect of IPTG Concentration on Recombinant Human Prethrombin-2 Expression inEscherichia coliBL21(DE3) ArcticExpress

Cited by 18 publications

References 19 publications

Effective Methods of Pyridoxine Supplementation in Laying Hens to Albumin and Globulin Levels

Effective Methods of Pyridoxine Supplementation in Laying Hens to Albumin and Globulin Levels

Display of lead‐binding proteins on Escherichia coli surface for lead bioremediation

Expression of human epidermal growth factor using Ssp DnaB mini-intein as fusion partner in Escherichia coli BL21(DE3)

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