• Korean Journal of Clinical Laboratory Science
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• v.47 no.1
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• pp.6-10
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• 2015

Photodynamic therapy (PDT) apply photosensitizers and light. The purpose of this study was to evaluate the in vitro efficacy of PDT using blue LED (light emitting diode) with photofrin and radachlorin for Propionibacterium acnes. The colony forming units method was used to assess the antibacterial activity. Suspension (1 mL) containing P. acnes at $1{\times}10^5CFU/mL$ were prepared and then 2 fold serial diluted to $12.5{\mu}g/mL$ from $50{\mu}g/mL$ concentration of photofrin and radachlorin. After 60 minutes incubation, light was irradiated for 10 to 30 minutes using the following light source of wavelength 460 nm, each energy density 36, 72 and $108J/cm^2$. Bacterial growth was evaluated after 72 hours incubation in a Phenylethanol Blood Agar (PEBA) culture. In addition, flow cytometric analysis were performed to measure the live cell after PDT. Also transmission electron microscopy (TEM) was employed to evaluate the effect of pathogens by PDT. The PDT Group was perfectly killed to all kind of photosensitizers dose of $12.5{\mu}g/mL$ with irradiation of 10 minutes. Also other Groups were killed to all kind of photosensitizers dose of $6.25{\mu}g/mL$ with irradiation time of 20 and 30 minutes. The flow cytometry showed a lower number of viable bacteria in the PDT group compared to the control group. The images of the TEM results were showed in cytoplasmic membrane damage and partially deformed to cell morphologies. These results suggest that radachlorin and photofrin combine blue LED PDT can be effectively treated when was proved treatment for acnes therapy.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1054-1059
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• 2005

An extracellular protease was identified from Bacillus subtilis KCCM 10257 by N-terminal sequencing and mass spectral analysis. The molecular mass of the extracellular protease was estimated to be 28 kDa by SDS-PAGE. Sequencing of the N-terminal of the protease revealed the sequence of A(G,S,R)QXVPYG(A)V(P,L)SQ. The N-terminal sequence exhibited close similarity to the sequence of other proteases from Bacillus sp. A mass list of the monoisotopic peaks in the MALDI-TOF spectrum was searched after peptide fragmentation of the protease. Six peptide sequences exhibiting monoisotopic masses of 1,276.61, 1,513.67, 1,652.81, 1,661.83, 1,252.61, and 1,033.46 were observed from the fragmented protease. These monisotopic masses corresponded to the lytic enzyme L27 from Bacillus subtilis 168, and the Mowse score was found to be 75. A doubly charged Top product (MS) at a m/z of 517.3 exhibiting a molecular mass of 1034.6 was further analyzed by de novo sequencing using a PE Sciex QSTAR Hybrid Quadropole-TOF (MS/MS) mass spectrometer. MS/MS spectra of the Top product (MS) at a m/z of 517.3 obtained from the fragmented peptide mixture of protease with Q-star contained the b-ion series of 114.2, 171.2, 286.2, 357.2, 504.2, 667.4, 830.1, and 887.1 and y-ion series of 147.5, 204.2, 367.2, 530.3, 677.4, 748.4, 863.4, and 920.5. The sequence of analyzed peptide ion was identified as LGDAFYYG from the b- and y-ion series by de novo sequencing and corresponded to the results from the MALDI-TOF spectrum. From these results the extracellular protease from Bacillus subtilis KCCM 10257 was successfully identified with the lytic enzyme L27 from Bacillus subtilis 168.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1568-1577
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• 2015

The purpose of this study was to investigate lactic acid bacteria with antioxidative and probiotic activities isolated from Korean healthy infant feces and kimchi. Isolates A1, A2, S1, S2, and S3 were assigned to Lactobacillus sp. and isolates A3, A4, E1, E2, E3, and E4 were assigned to Leuconostoc sp. on the basis of their physiological properties and 16S ribosomal DNA sequence analysis. Most strains were confirmed as safe bioresources through nonhemolytic activities and non-production of harmful enzymes such as β-glucosidase, β-glucuronidase and tryptophanase. The 11 isolates showed different resistance to acid and bile acids. In addition, they exhibited antibacterial activity against foodborne bacteria, especially Bacillus cereus, Listeria monocytogenes, and Escherichia coli. Furthermore, all strains showed significantly high levels of hydrophobicity. The antioxidant effects of culture filtrates of the 11 strains included 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging capacity, 2.2'-azino-bis (2-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation scavenging activity, and superoxide dismutase activity. The results revealed that most of the culture filtrates have effective scavenging activity for DPPH and ABTS radicals. All strains appeared to have effective superoxide dismutase activity. In conclusion, the isolated strains A1, A3, S1, and S3 have significant probiotic activities applicable to the development of functional foods and health-related products. These strains might also contribute to preventing and controlling several diseases associated with oxidative stress, when used as probiotics.

• Journal of Microbiology and Biotechnology
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• v.21 no.12
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• pp.1330-1335
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• 2011

N-Acyl homoserine lactones (AHLs) serve as the vital quorum-sensing signals that regulate the virulence of the pathogenic bacterium Erwinia carotovora. In the present study, an approach to efficiently restrain the pathogenicity of E. carotovora-induced soft rot disease is described. Bacillus thuringiensis-derived N-acyl homoserine lactonase (AiiA) was projected onto the surface of Pseudomonas putida cells, and inoculation with both strains was challenged. The previously identified N-terminal moiety of the ice nucleation protein, InaQ-N, was applied as the anchoring motif. A surface display cassette with inaQ-N/aiiA was constructed and expressed under the control of a constitutive promoter in P. putida AB92019. Surface localization of the fusion protein was confirmed by Western blot analysis, flow cytometry, and immunofluorescence microscopy. The antagonistic activity of P. putida MB116 expressing InaQ-N/AiiA toward E. carotovora ATCC25270 was evaluated by challenge inoculation in potato slices at different ratios. The results revealed a remarkable suppressing effect on E. carotovora infection. The active component was further analyzed using different cell fractions, and the cell surface-projected fusion protein was found to correspond to the suppressing effect.

• Journal of Microbiology and Biotechnology
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• v.28 no.11
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• pp.1814-1822
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• 2018

Most people use cosmetics to protect their skin. Preservatives are often used to prevent their contamination upon use. There has been a great demand for natural preservatives due to recent reports on the side effects of parabens. Therefore, we evaluated the antimicrobial activities of Lonicera japonica and Magnolia obovata extracts and determined their potential as natural preservatives. We found that the 50% ethanol extract from L. japonica had antibacterial activity only against S. aureus and P. aeruginosa, while the ethyl acetate fraction showed antimicrobial activity against all six microbial strains tested. On the other hand, the 70% ethanol extract and the ethyl acetate fraction from M. obovata showed antimicrobial activity against all six strains. A synergistic effect against S. aureus, B. subtilis, and C. albicans was confirmed when two ethyl acetate fractions having antimicrobial activity against all six strains were used in combination. Synergistic activity against B. subtilis was also confirmed through kill-time analysis. High-performance liquid chromatography was performed to identify the components of each extract. Based on the minimum inhibitory concentration and the results of a disc diffusion assay, we confirmed that caffeic acid and luteolin influenced the antimicrobial activity of L. japonica and that the antimicrobial activity of M. obovata was influenced by the interaction of magnolol and honokiol with other components. Therefore, this study suggests that the combination of L. japonica and M. obovata extracts may be used as a plant-derived natural preservative.

• Journal of Microbiology and Biotechnology
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• v.23 no.10
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• pp.1381-1385
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• 2013

The centipede Scolopendra subpinipes mutilans is a medicinally important arthropod species. However, its transcriptome is not currently available and transcriptome analysis would be useful in providing insight into a molecular level approach. Hence, we performed de novo RNA sequencing of S. subpinipes mutilans using next-generation sequencing. We generated a novel peptide (scolopendrasin II) based on a SVM algorithm, and biochemically evaluated the in vitro antimicrobial activity of scolopendrasin II against various microbes. Scolopendrasin II showed antibacterial activities against gram-positive and -negative bacterial strains, including the yeast Candida albicans and antibiotic-resistant gram-negative bacteria, as determined by a radial diffusion assay and colony count assay without hemolytic activity. In addition, we confirmed that scolopendrasin II bound to the surface of bacteria through a specific interaction with lipoteichoic acid and a lipopolysaccharide, which was one of the bacterial cell-wall components. In conclusion, our results suggest that scolopendrasin II may be useful for developing peptide antibiotics.

• KSBB Journal
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• v.31 no.4
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• pp.228-236
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• 2016

The aim of this study was to characterize the hemolytic Aeromonas sp. MH-8 exposed to green tea catechin, epigallocatechin gallate (EGCG). Initially, the hemolytic Aeromonas sp. MH-8 was enriched and isolated from stale fish. Bactericidal effects of MH-8 exposed to EGCG ranging from 1 mg/mL to 4 mg/mL were monitored, and complete bactericidal effects were achieved within 3 h at 3 mg/mL and higher concentrations. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain MH-8 treated to different concentrations and exposing periods of EGCG in exponentially growing cultures. The stress shock proteins (70-kDa DnaK and 60-kDa GroEL), which might contribute to enhancing the cellular resistance to the cytotoxic effect of EGCG, were induced at different concentrations of EGCG exposed to cell culture of MH-8. Scanning electron microscopic analysis demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with EGCG. 2-DE of soluble protein fractions from MH-8 cultures showed 18 protein spots changed by EGCG exposure. These proteins involved in chaperons (e.g., DnaK, GroEL and trigger factor), enterotoxins (e.g., aerolysin and phospholipase C precursor), LPS synthesis (e.g., LPS biosynthesis protein and outer membrane protein A precursor), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. In consequence, EGCG was found to have substantial antibacterial effects against food-poisoning causing bacterium, hemolytic Aeromonas sp. MH-8. Also the results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on Aeromonas sp. MH-8.

• Fashion & Textile Research Journal
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• v.18 no.4
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• pp.531-539
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• 2016

Charcoal dyed fabrics have been widely used in home textiles and functional clothing due to their anti-statics, antibacterial, deodorization, far infrared emitting and anion releasing. Soybean fiber were regenerated from soybean. Soybean fiber have biodegradable, microbiocidal, non-allergic, and anti-ageing properties. The purpose of this study is to investigate the dyeing characteristics of soybean fabric using charcoal as colorants. Soybean fabrics were dyed with charcoal solution according to concentration of charcoal, dyeing temperature, and dyeing time. To improve washing fastness and investigate mordanting condition, soybean fabric and dyed soybean fabric with charcoal were mordanted by mordanting agents such as $CH_3COOH$(acetic acid), NaCl(sodium chloride) and $AlK(SO_4)_2{\cdot}12H_2O$(Aluminium Potassium Sulfate). Dyeability and color characteristics of charcoal dyed soybean fabric were obtained by computer color matching and SEM morphology analysis. Particle size of charcoal and color fastness were also investigated. The results obtained were as follows; Mean average diameter of charcoal was $1.39{\mu}m$. The dyeability of soybean fabric using charcoal as colorants was increased gradually with increasing concentration of charcoal dyeing solution and saturated at about 8%(o.w.b.). The optimum dyeing temperature and dyeing time were $90{\sim}105^{\circ}C$ and 60~90 minutes respectively. The overall wash fastness at dyeing concentration 2~4%(o.w.b.) and 6~10%(o.w.b.) were 4 degree and 3-4 degree respectively. The fastness to washing according to mordanting method indicated good grade result as more than 4 degree in all conditions. On the other hand, the staining of adjacent fabrics, i.e. PET, Acryl, Wool, Acetate, Nylon and Cotton was found to be of grade 4 or 4-5 in all conditions.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.8
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• pp.1132-1136
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• 2010

After Cheonnyuncho (Opuntia humifusa) was extracted by 70% ethanol and partitioned with hexane, chloroform, ethyl acetate, butanol, and water in order, ethyl acetate fraction with excellent antioxidant activities was acquired. Ethyl acetate fraction was conducted by TLC, and the third spot of 10 spots was selected due to its best antioxidant activities. When the third fraction further isolated on silica gel chromatography, the fourth fraction of 10 fractions had the best antioxidant activities and purified by HPLC. The molecular weight on EI-MS, and $^1H$- and $^{13}C$-NMR spectrum showed that the purified compound was taxifolin. In addition, antioxidant activities were analyzed, and the purified compound from Cheonnyuncho was found to be effective as much as $\alpha$-tocopherol, BHA. According to the analysis on antibacterial activities, the purified compound also showed more activity than benzoic acid against all pathogenic bacteria.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.442-448
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• 2006

Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.