Purification of plant complex protein extracts in non-denaturing conditions by in-solution isoelectric focusing

Ferreira, Renata; Dias, Sandra

doi:10.1016/j.ab.2016.07.004

Cited by 6 publications

(3 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Complexes separated by non‐denaturing IEF can be eluted from the cathode end of the IEF gel. It has been reported that native proteins and protein complexes in solution can be separated and extracted by offgel IEF, because a maximum current of 50 μA per IEF and 200 mW of power was used in this method . In the present study, a current of 0.1 mA per non‐denaturing IEF gel column was applied, and less than 150 mW of power was used.…”

Section: Resultsmentioning

confidence: 92%

Microscale isolation of native forms of lysozyme from chicken egg white by gel isoelectric focusing

2018

View full text Add to dashboard Cite

To separate and extract the native states of lysozyme from chicken egg white, a hybrid method for the mobilization of proteins after non-denaturing gel isoelectric focusing (IEF) combined with detection of lysozyme activity was developed. When the proteins in the chicken egg white were first separated using non-denaturing gel IEF, a lysozyme was obtained at the top of the gel column at the cathode end of the IEF. And, when the IEF-separated proteins of the egg white were mobilized by replacing the cathodic sodium hydroxide solution with phosphoric acid solution, an additional active state of the lysozyme that could be bound to proteins, such as ovotransferrin, was extracted from the solution. Furthermore, it was shown that the addition of lysozyme, obtained via IEF, to pure ovotransferrin generated a complex manifesting lysozyme activity, clearly indicating a successful reconstruction of the lysozyme-ovotransferrin complex in vitro. Therefore, the obtained results demonstrated that the native states of lysozymes, such as lysozyme and the lysozyme-ovotransferrin complex, can be effectively separated and extracted using non-denaturing gel IEF. Thus, this method can be applied to separate and extract different charge states of native proteins that retain their biological activities.

show abstract

Section: Resultsmentioning

confidence: 92%

Microscale isolation of native forms of lysozyme from chicken egg white by gel isoelectric focusing

2018

View full text Add to dashboard Cite

show abstract

“…58 The samples were fractionated using a 24-well setup and the OFFGEL kit 3–10 (Agilent Technologies) and a 3100 OFFGEL Fractionator. Contrary to the manufacturer’s instructions, protein denaturing and reducing agents were not added to the OGE stock solution.…”

Section: Separation Modes and Conditionsmentioning

confidence: 99%

Recent advances in protein analysis by capillary and microchip electrophoresis

Dawod

Arvin

Kennedy

2017

Analyst

117

View full text Add to dashboard Cite

This review article describes the significant recent advances in analysis of proteins by capillary and microchip electrophoresis during the period mid 2014 to early 2017. The review highlights the progressions, new methodologies, innovative instrumental modifications, and challenges for efficient protein analysis in human specimens, animal tissues, and plant samples. Protein analysis fields covered in this review include analysis of native, reduced, and denatured proteins in addition to Western blotting, protein therapeutics and proteomics.

show abstract

“…Among the many alternatives, such as plant extracts, antimicrobial peptides, and bee products, metal or metal oxide nanoparticles are recently receiving increased attention [19][20][21][22]. The properties found in the nanoparticle form of these materials can signi cantly differ from those of their bulk tantamount.…”

Section: Introductionmentioning

confidence: 99%

Precise engineering of ultrasound-mediated cerium oxide nanoparticles: Investigations on antioxidant, antibacterial activity and human osteomyelitis

Dai

Wang

Chen

et al. 2021

Preprint

View full text Add to dashboard Cite

In this study, the antibacterial activity of cerium oxide nanoparticles on two Gram-negative and three Gram-positive foodborne pathogens was investigated. CeO2 nanoparticles (CeO2 NPs) were synthesized by a Wet Chemical Synthesis route, using the precipitation method and the Simultaneous Addition of reactants (WCS–SimAdd). The as-obtained precursor powders were investigated by thermal analysis (TG–DTA), to study their decomposition process and to understand the CeO2 NPs formation. The composition, structure, and morphology of the thermally treated sample were investigated by FTIR, Raman spectroscopy, X-ray diffraction, TEM, and DLS. The cubic structure and average particle size ranging between 5 and 15 nm were evidenced. Optical absorption measurements (UV–Vis) reveal that the band gap of CeO2 NPs is 2.61 eV, which is smaller than the band gap of bulk ceria. The antioxidant effect of CeO2 NPs was determined, and the antibacterial test was carried out both in liquid and on solid growth media against five pathogenic microorganisms, namely Escherichia coli, Salmonella typhimurium, Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus. Cerium oxide nanoparticles showed growth inhibition toward all five pathogens tested with notable results. This paper highlights the CeO2 NPs showed antibacterial activity with significant variations due to the differences in the membrane structure and cell wall composition among the two groups tested. Consequently, synthesized CeO2 NPs can be potential candidates for the treatment of osteomyelitis.

show abstract

Purification of plant complex protein extracts in non-denaturing conditions by in-solution isoelectric focusing

Cited by 6 publications

References 16 publications

Microscale isolation of native forms of lysozyme from chicken egg white by gel isoelectric focusing

Microscale isolation of native forms of lysozyme from chicken egg white by gel isoelectric focusing

Recent advances in protein analysis by capillary and microchip electrophoresis

Precise engineering of ultrasound-mediated cerium oxide nanoparticles: Investigations on antioxidant, antibacterial activity and human osteomyelitis

Contact Info

Product

Resources

About