Protein profile characterization of bacterial lysates by capillary electrophoresis

Kustos, Ildikó; Kocsis, Béla; Kerepesi, Ildikó; Kilár, Ferenc

doi:10.1002/elps.1150191311

Cited by 28 publications

(21 citation statements)

References 23 publications

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“…The bacterial protein profiles are a reflection of the genome of the strain, and, therefore, determination of the whole protein content plays an important role in the comparative studies of bacteria (25). To further confirm the permeabilizing action of sericin, E. coli cells were analyzed for soluble proteins by SDS-PAGE.…”

Section: Resultsmentioning

confidence: 99%

Shape Changes and Interaction Mechanism of Escherichia coli Cells Treated with Sericin and Use of a Sericin-Based Hydrogel for Wound Healing

Xue

Liu

Zhang

et al. 2016

Appl Environ Microbiol

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To verify the interaction mechanism between sericin and Escherichia coli, especially the morphological and structural changes in the bacterial cells, the antimicrobial activity of sericin against E. coli as a model for Gram-negative bacteria was investigated. The antibacterial activity of sericin on E. coli and the interaction mechanism were investigated in this study by analyzing the growth, integrity, and morphology of the bacterial cells following treatment with sericin. The changes in morphology and cellular compositions of bacterial cells treated with sericin were observed by an inverted fluorescence microscope, scanning electron microscopy, and transmission electron microscopy. Changes in electrical conductivity, total sugar concentration of the broth for the bacteria, and protein expression of the bacteria were determined to investigate the permeability of the cell membrane. A sericinbased hydrogel was prepared for an in vivo study of wound dressing. The results showed that the antibacterial activity of the hydrogel increased with the increase in the concentration of sericin from 10 g/liter to 40 g/liter. The introduction of sericin induces membrane blebbing of E. coli cells caused by antibiotic action on the cell membrane. The cytoplasm shrinkage phenomenon was accompanied by blurring of the membrane wall boundaries. When E. coli cells were treated with sericin, release of intracellular components quickly increased. The electrical conductivity assay indicated that the charged ions are reduced after exposure to sericin so that the integrity of the cell membrane is weakened and metabolism is blocked. In addition, sodium dodecyl sulfatepolyacrylamide gel electrophoresis demonstrated that sericin hinders the expression of bacterial protein. Sericin may damage the integrity of the bacterial cell membrane, thereby eventually inhibiting the growth and reproduction of E. coli. Compared to sterile gauze, the sericin-based hydrogel promoted fibroblast cell proliferation and accelerated the formation of granulation tissues and neovessels. IMPORTANCEThe specific relationship and interaction mechanism between sericin and E. coli cells were investigated and elucidated. The results show that after 12 h of treatment, sericin molecules induce membrane blebbing of E. coli cells, and the bacteria show decreases in liquidity and permeability of biological membrane, resulting in alterations in the conductivity of the culture medium and the integrity of the outer membrane. The subsequent in vivo results demonstrate that the sericin-poly(N-isopropylacrylamide-N,N=-methylene-bis-acrylamide [NIPAm-MBA]) hydrogel accelerated wound healing compared to that with sterile gauze, which is a beneficial result for future applications in clinical medicine and the textile, food, and coating industries.A lthough many bacteria, such as bifidobacteria and lactobacilli, are beneficial for our daily lives, harmful bacteria like Staphylococcus aureus and Escherichia coli are serious threats to human health due to rapid proliferation and...

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Section: Resultsmentioning

confidence: 99%

Shape Changes and Interaction Mechanism of Escherichia coli Cells Treated with Sericin and Use of a Sericin-Based Hydrogel for Wound Healing

Xue

Liu

Zhang

et al. 2016

Appl Environ Microbiol

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“…The bacterial protein profiles are a reflection of the genome of the strain; therefore, determination of the whole protein content plays an important role in classification, identification, typing, and comparative (Kustos et al, 1998). SDS-PAGE is also an important molecular technique used for identification at the species level (Durrani et al, 2008).…”

Section: Resultsmentioning

confidence: 99%

Analysis of whole cell protein profiles of Salmonella serovars isolated from chicken, turkey and sheep faeces by SDS-PAGE

Aksakal¹

2010

Vet. Med.

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This study was carried out to determine the whole cell protein profiles of Salmonella serovars from chicken, turkey and sheep faeces by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A total of 34 Salmonella strains were included in the study, 14 of them were isolated from chicken, 14 from turkey and six from sheep. SDS-PAGE was carried out using 12% (w/v) separating and 4% (w/v) stacking gels. The results showed more than 30 protein bands ranging in size from 97 kDa (kilodaltons) to below 14.4 kDa as determined by visual assessment of their approximate molecular masses. Protein bands of 78.1, 51.2, 41.5, 37.3, 35.1, 33.9, 30.7, 27.6, 25.4, and 24 kDa were detected in all Salmonella serovars. Salmonella strains used in this study were closely related and could not be differentiated depending on the whole cell protein profiles using SDS-PAGE.

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“…In the SDS-CGE method, a high viscosity polymer is used as a buffer [16,[23][24][25][26][27][28]. Though sharp peaks were obtained, the high viscosity of the buffer decreased the efficiency of the injection, preventing microchannel injection.…”

Section: Design Of the Hts Systems For Proteome Analysismentioning

confidence: 99%

A separation carrier in high-speed proteome analysis by capillary electrophoresis

Tabuchi

Baba

2001

Electrophoresis

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We obtained a high-efficiency separation carrier for proteome analysis by capillary electrophoresis. The addition of curdlan or laminaran to the run buffer hastened the migration time without any degradation in resolution. We propose that for the development of the separation carrier it is necessary to synthetically analyze each of the following mobility factors of electroosmotic flow: buffer ionic strength, additional disturbance and adsorption. The total analysis for buffer and additive will be useful for designing high-throughput screening (HTS) systems for proteome analysis without annoying adsorption.

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Protein profile characterization of bacterial lysates by capillary electrophoresis

Cited by 28 publications

References 23 publications

Shape Changes and Interaction Mechanism of Escherichia coli Cells Treated with Sericin and Use of a Sericin-Based Hydrogel for Wound Healing

Shape Changes and Interaction Mechanism of Escherichia coli Cells Treated with Sericin and Use of a Sericin-Based Hydrogel for Wound Healing

Analysis of whole cell protein profiles of Salmonella serovars isolated from chicken, turkey and sheep faeces by SDS-PAGE

A separation carrier in high-speed proteome analysis by capillary electrophoresis

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