Evaluation of immobilized metal affinity chromatography kits for the purification of histidine-tagged recombinant CagA protein

Karakus, Cebrail; Uslu, Merve; Yazıcı, Duygu; Salih, Barik A.

doi:10.1016/j.jchromb.2015.11.045

Cited by 19 publications

(10 citation statements)

References 13 publications

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“…In the mid-1970s, Porath et al first described immobilized metal ion affinity chromatography (IMAC) [11]. Since that time, proteins of interest have been enriched through IMAC [12,13,14,15,16]. Nickel ion immobilized on solid supports have been widely used in IMAC [17,18].…”

Section: Introductionmentioning

confidence: 99%

Separation and Characterization of Angiotensin I Converting Enzyme (ACE) Inhibitory Peptides from Saurida elongata Proteins Hydrolysate by IMAC-Ni2+

Sun

Zhou

et al. 2017

Marine Drugs

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Lizard fish protein hydrolysates (LFPH) were prepared from Lizard fish (Saurida elongata) proteins possessing powerful angiotensin I converting enzyme (ACE) inhibitory activity and the fraction (LFPH-I) with high ACE inhibitory activity was obtained through ultrafiltration. The active Fraction (F2) was isolated from LFPH-I using immobilized metal affinity chromatography (IMAC-Ni2+). Analysis of amino acid levels revealed that F2 eluted from IMAC was enriched in Met, His, Tyr, Pro, Ile, and Leu compared to the crude peptide LFPH-I. F2 with the high ACE inhibitory activity (IC50 of 0.116 mg·mL−1) was further separated by a reverse-phase column to yield a novel ACE inhibitory peptide with IC50 value of 52 μM. The ACE inhibitory peptide was identified as Arg-Tyr-Arg-Pro, RYRP. The present study demonstrated that IMAC may be a useful tool for the separation of ACE inhibitory peptides from protein hydrolysate.

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Section: Introductionmentioning

confidence: 99%

Separation and Characterization of Angiotensin I Converting Enzyme (ACE) Inhibitory Peptides from Saurida elongata Proteins Hydrolysate by IMAC-Ni2+

Sun

Zhou

et al. 2017

Marine Drugs

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“…Cultivation of buwchitin transformants were scaled up to a total volume of 1 L to produce cell pellets for protein purification. Recombinant proteins were purified in their native conditions to preserve their activity (Karakus et al, 2016). Figure 1 shows the SDS-PAGE analysis of the purification fractions for buwchitin.…”

Section: Resultsmentioning

confidence: 99%

Buwchitin: A Ruminal Peptide with Antimicrobial Potential against Enterococcus faecalis

et al. 2017

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Antimicrobial peptides (AMPs) are gaining popularity as alternatives for treatment of bacterial infections and recent advances in omics technologies provide new platforms for AMP discovery. We sought to determine the antibacterial activity of a novel antimicrobial peptide, buwchitin, against Enterococcus faecalis. Buwchitin was identified from a rumen bacterial metagenome library, cloned, expressed and purified. The antimicrobial activity of the recombinant peptide was assessed using a broth microdilution susceptibility assay to determine the peptide's killing kinetics against selected bacterial strains. The killing mechanism of buwchitin was investigated further by monitoring its ability to cause membrane depolarization (diSC3(5) method) and morphological changes in E. faecalis cells. Transmission electron micrographs of buwchitin treated E. faecalis cells showed intact outer membranes with blebbing, but no major damaging effects and cell morphology changes. Buwchitin had negligible cytotoxicity against defibrinated sheep erythrocytes. Although no significant membrane leakage and depolarization was observed, buwchitin at minimum inhibitory concentration (MIC) was bacteriostatic against E. faecalis cells and inhibited growth in vitro by 70% when compared to untreated cells. These findings suggest that buwchitin, a rumen derived peptide, has potential for antimicrobial activity against E. faecalis.

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“…Metal ions interact with nitrogen atoms locating within the structure of amino acids (especially nitrogen atoms in the imidazole ring of histidine amino acids) on the surface of proteins (20).…”

Section: Immobilized Metal Affinity Chromatography (Imac) Has Been Dementioning

confidence: 99%

The Adsorption of Calmoduline via Nicotinamide Immobilized Poly(HEMA-GMA) Cryogels

Erol¹

2016

Journal of the Turkish Chemical Society, Section A: Chemistry

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Abstract:The separation and purification of an important biomolecule calmoduline is extremely important. The adsorption technique is quite popular, and the cryogels as adsorbents with the macro porous structure and interconnected flow channels are preferred in this field. In this study, the adsorption of calmoduline via Ca(II) immobilized poly(2-hydroxyethyl methacrylate-glycidyl methacrylate), poly(HEMA-GMA), cryogels was studied through changing interaction time, calmoduline initial concentration, and temperature conditions. For the characterization of cryogels, the swelling test, Fourier Transform Infrared (FT-IR) Spectroscopy, Scanning Electron Microscopy (SEM), surface area (BET), elemental analysis and ICP-OES methods were performed. Nicotinamide molecule was used as Ca(II) being the chelating agent and the adsorption capacity of the cryogels was estimated as 1.812 mg calmoduline/g cryogel. The adsorption models of the process were examined via the Langmuir and Freundlich isotherm models.

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Evaluation of immobilized metal affinity chromatography kits for the purification of histidine-tagged recombinant CagA protein

Cited by 19 publications

References 13 publications

Separation and Characterization of Angiotensin I Converting Enzyme (ACE) Inhibitory Peptides from Saurida elongata Proteins Hydrolysate by IMAC-Ni2+

Separation and Characterization of Angiotensin I Converting Enzyme (ACE) Inhibitory Peptides from Saurida elongata Proteins Hydrolysate by IMAC-Ni2+

Buwchitin: A Ruminal Peptide with Antimicrobial Potential against Enterococcus faecalis

The Adsorption of Calmoduline via Nicotinamide Immobilized Poly(HEMA-GMA) Cryogels

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