• Title/Summary/Keyword: antimicrobial substrate

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Enzymatic Hydrolysis of Ovotransferrin and the Functional Properties of Its Hydrolysates

  • Rathnapala, Ethige Chathura Nishshanka;Ahn, Dong Uk;Abeyrathne, Edirisingha Dewage Nalaka Sandun
    • Food Science of Animal Resources
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    • v.41 no.4
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    • pp.608-622
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    • 2021
  • Bioactive peptides have great potentials as nutraceutical and pharmaceutical agents that can improve human health. The objectives of this research were to produce functional peptides from ovotransferrin, a major egg white protein, using single enzyme treatments, and to analyze the properties of the hydrolysates produced. Lyophilized ovotransferrin was dissolved in distilled water at 20 mg/mL, treated with protease, elastase, papain, trypsin, or α-chymotrypsin at 1% (w/v) level of substrate, and incubated for 0-24 h at the optimal temperature of each enzyme (protease 55℃, papain 37℃, elastase 25℃, trypsin 37℃, α-chymotrypsin 37℃). The hydrolysates were tested for antioxidant, metal-chelating, and antimicrobial activities. Protease, papain, trypsin, and α-chymotrypsin hydrolyzed ovotransferrin relatively well after 3 h of incubation, but it took 24 h with elastase to reach a similar degree of hydrolysis. The hydrolysates obtained after 3 h of incubation with protease, papain, trypsin, α-chymotrypsin, and after 24 h with elastase were selected as the best products to analyze their functional properties. None of the hydrolysates exhibited antioxidant properties in the oil emulsion nor antimicrobial property at 20 mg/mL concentration. However, ovotransferrin with α-chymotrypsin and with elastase had higher Fe3+-chelating activities (1.06±0.88%, 1.25±0.24%) than the native ovotransferrin (0.46±0.60%). Overall, the results indicated that the single-enzyme treatments of ovotransferrin were not effective to produce peptides with antioxidant, antimicrobial, or Fe3+-chelating activity. Further research on the effects of enzyme combinations may be needed.

Antimicrobial and Antioxidative Activities of Solvent Fraction from Humulus japonicus (환삼덩굴의 용매분획별 항균성 및 항산화성)

  • Park, Seung-Woo;Woo, Cheol-Joo;Chung, Shin-Kyo;Chung, Ki-Taek
    • Korean Journal of Food Science and Technology
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    • v.26 no.4
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    • pp.464-470
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    • 1994
  • The biological activities of Humulus japonicus were extracted by water and methanol. Methanol was better solvent than water in the extraction for antimicrobial activities against six different species of bacteria and two yeasts. The methanol extract was systematically fractionated with various organic solvents which have different polarities. From the result of antimicrobial activities against six species of bacteria and two species of yeasts, methanol extract was superior to water extract. The methanol extract of Humulus japonicus showed antimicrobial activity against the all species of microorganisms tested except Escherichia coli . The butanol fraction of methanol extract showed antimicrobial effect on the all species tested. The minimal inhibition concentration(MIC) of the butanol fraction on the growth of microorganisms was ranged between $0.1{\sim}0.4%$. The water extract of Humulus japonicus did not show inhibition of the activity of trypsin but methanol extract showed inhibitory activity. The chloroform fraction of methanol extract showed comparatively higher trypsin inhibitory activity than other fractions. The concentration of 50% inhibition$(IC_{50})$ by chloroform fraction was 1.0 mg/ml. Enzyme-inhibitor complex formation was above 90% of the while for 20 min. It was revealed that methanol extract of Humulus japonicus inhibited peroxide production of lard and soybean oil as substrate by antioxidative test. The chloroform fraction of methanol extract had the highest activity. When 0.2% of chloroform fraction was added, induction period of soybean oil and lard were extended 15, 9 days, respectively.

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Numerical Classification of Actinomycetes Isolated from Volcanic Soil

  • Kim, Seung-Bum;Lee, Soon-Dong;Kim, Seon-Young;Oh, Hyung-Myung;Kang, Sa-Ouk
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.105-116
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    • 1996
  • Of actinomycetes isolated from volcanic compost soils, 115 representative strains which showed distinctive morphologicla features were numerically classified, compared with reference strains of Streptomyces. One hundred and twenty unit characters were tested and the average probability of error was 4.27%. The cluster analysis resulted in two groups: group A included strains of actinomycetes except streptomycetes. Group A was divided into 2 major clusters (over 5 strains), 10-diaminopimelic acid. Group B was divided into 5 clusters, of which 4 clusters contained mesodiminopimelic acid and 1 cluster LL-diaminopimelic acid. The major clusters of group A showed higher abilities of substrate utilization and degradation, and higher resistance to inhibitors, whereas the minor and single member clusters of group A showed relatively higher antimicrobial activities. On the other hand, all clusters of group B showed relatively lower abilities of substrate utilization and degradation and lower resistance to inhibitors.

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Influence of Silver Ions in HA Film on Morphology of Macrophages

  • Feng, Q.L.;Kim, T.N.;Kim, J.W.
    • Journal of the Korean Vacuum Society
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    • v.7 no.s1
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    • pp.50-56
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    • 1998
  • Ion beam assisted deposition (IBAD) was successfully used to produce a dense and ultra-adherent Hydroxyapatite (HA) film on titanium alloy and alumina. Recently it is also proved that the HA coatings on alumina substrate treated with 20 ppm $AgNO_3$ had the structure of $(Ag, Ca)_10(PO_4)6(OH)_2$, which exhibited excellent antimicrobial effects. The present paper aims to morphlogically characterize the adhesion of macrophages on newly developed Ag-HA coated alumina and Ti6A14V substrates and to evaluate the biocompatibility of the coatings in vitro. It can be found that the cell number on alumina of the concentration of $AgNO_3$ in the treatment, the cell number on Ag-HA coatings decreased. Up to 20 ppm $AgNO_3$ by Ag-treatment, the morphological development of the cells on Ag-HA coating was similar to that of the cells on HA coating, suggesting the biotolerance of the Ag-HA coatings.

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Effect of Supplementation of Bacillus subtilis LS 1-2 Grown on Citrus-juice Waste and Corn-soybean Meal Substrate on Growth Performance, Nutrient Retention, Caecal Microbiology and Small Intestinal Morphology of Broilers

  • Sen, Sinol;Ingale, S.L.;Kim, J.S.;Kim, K.H.;Kim, Y.W.;Khong, Chou;Lohakare, J.D.;Kim, E.K.;Kim, H.S.;Kwon, I.K.;Chae, B.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.8
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    • pp.1120-1127
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    • 2011
  • A feeding trial was conducted to investigate the effect of dietary supplementation of Bacillus subtilis LS 1-2 grown on citrus-juice waste and corn-soybean substrate on growth performance, nutrient retention, caecal microbial population and intestinal morphology in broilers. Three hundred twenty d-old Ross chicks were randomly allotted to 4 treatments on the basis of BW in a randomized complete block design. Each treatment had 4 replicates of 20 chicks in each. Experimental diets were fed in 2 phases, starter (d 0 to 21) and finisher (d 21 to 35). Dietary treatments were; negative control (NC: basal diet without any antimicrobial), positive control (PC: basal diet added with 20 mg/kg Avilamycin), basal diet added with 0.30% Bacillus subtilis LS 1-2 grown on corn-soybean substrate (P1), and basal diet added with 0.30% Bacillus subtilis LS 1-2 grown on citrus-juice waste substrate (P2). Overall BW gain, feed intake and FCR were better (p<0.05) in PC, P1 and P2 treatments as compared to NC. Moreover, overall BW gain and FCR in PC and P2 treatments were greater than P1. Retention of CP, GE (d 21, d 35) and DM (d 35) were increased (p<0.05) in treatments PC, P1 and P2 compared with NC. At d 35, caecal Clostridium and Coliform counts were lower (p<0.05) in treatments PC, P1 and P2 than NC. Moreover, Clostridium and Coliform counts in treatment PC was lower (p<0.05) than P1. Villus height and villus height to crypt depth ratio in both duodenum and ileum were increased (p<0.05) in treatments PC, P1, P2 as compared to NC. However, retention of nutrients, caecal microbial population and intestinal morphology remained comparable among treatments P1 and P2. It is concluded that Bacillus subtilis LS 1-2 inclusion at 0.30% level had beneficial effects on broilers' growth performance, nutrient retention, caecal microflora and intestinal morphology. Additionally, citrus-juice waste can be used as substrate for growth of probiotic Bacillus subtilis LS 1-2.

Prebiotic Potential of Xylooligosaccharides Derived from Corn Cobs and Their In Vitro Antioxidant Activity When Combined with Lactobacillus

  • Yu, Xiuhua;Yin, Jianyuan;Li, Lin;Luan, Chang;Zhang, Jian;Zhao, Chunfang;Li, Shengyu
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1084-1092
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    • 2015
  • In the present work, the in vitro prebiotic activity of xylooligosaccharides (XOS) derived from corn cobs combined with Lactobacillus plantarum, a probiotic microorganism, was determined. These probiotics exhibited different growth characteristics depending on strain specificity. L. plantarum S2 cells were denser and their growth rates were higher when cultured on XOS. Acetate was found to be the major short-chain fatty acid produced as the end-product of fermentation, and its amount varied from 1.50 to 1.78 mg/ml. The antimicrobial activity of XOS combined with L. plantarum S2 was determined against gastrointestinal pathogens. The results showed that XOS proved to be an effective substrate, enhancing antimicrobial activity for L. plantarum S2. In vivo evaluation of the influence of XOS and L. plantarum S2, used both alone and together, on the intestinal microbiota in a mouse model showed that XOS combined with L. plantarum S2 could increase the viable lactobacilli and bifidobacteria in mice feces and decrease the viable Enterococcus, Enterobacter, and Clostridia spp. Furthermore, in the in vitro antioxidant assay, XOS combined with L. plantarum S2 possessed significant 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis, and superoxide anion radical-scavenging activities, and the combinations showed better antioxidant activity than either XOS or L. plantarum S2 alone.

Expression, Purification and Properties of Shikimate Dehydrogenase from Mycobacterium Tuberculosis

  • Zhang, Xuelian;Zhang, Shunbao;Hao, Fang;Lai, Xuhui;Yu, Haidong;Huang, Yishu;Wang, Honghai
    • BMB Reports
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    • v.38 no.5
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    • pp.624-631
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    • 2005
  • Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the main diseases to mankind. It is urgent to discover novel drug targets for appropriate antimicrobial agents against this human pathogen. The shikimate pathway is onsidered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammalian cells. The Mycobacterium tuberculosis aroE-encoded shikimate dehydrogenase was cloned, expressed and purified. Sequence alignment analysis shows that shikimate dehydrogenase of Mycobacterium tuberculosis exhibit the pattern of G-X-(N/S)-V-(T/S)-X-PX-K, which is highly conserved within the shikimate dehydrogenase family. The recombinant shikimate dehydrogenase spectrum determined by CD spectroscopy showed that the percentages for $\alpha$-helix, $\beta$-sheet, $\beta$-turn, and random coil were 29.2%, 9.3%, 32.7%, and 28.8%, respectively. The enzymatic characterization demonstrates that it appears to be fully active at pH from 9.0 to 12, and temperature $63^{\circ}C$. The apparent Michaelis constant for shikimic acid and $NADP^+$ were calculated to be about $29.5\;{\mu}M$ and $63\;{\mu}M$. The recombinant shikimate dehydrogenase catalyzes the substrate in the presence of $NADP^+$ with an enzyme turnover number of $399\;s^{-1}$. Zymological studies suggest that the cloned shikimate dehydrogenase from M. tuberculosis has a pretty activity, and the work should help in the discovery of enzyme inhibitors and further of possible antimicrobial agents against Mycobacterium tuberculosis.

Evaluation of biological activity for Dangyuja (Citrus grandis) leaves and investigation of optimal concentrations extracted by alternative ethanol concentrations (에탄올 농도별 당유자 잎의 최적추출조건 및 생리활성 평가)

  • Nakamura, Masaya;Ra, Jong-Hwan;Kim, Ju-Sung
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.45-55
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    • 2019
  • TheCitrus grandis Osbeck is a special product in the Jeju island. The product has been as a remedy for liver damage and hang over. This study demonstrates how to investigate and compare the antioxidant, phenol content, tyrosinase and ${\alpha}$-glucosidase inhibitory activity, antimicrobial, and alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity with the C. grandis leaves extracted in different ethanol concentrations. From the yield, a 20% ethanol extract demonstrated the highest results among the other extracts. The distilled water extract showed the most abundant in a total phenol content and highest ABTS radical scavenging activity and reducing power assay. In the DPPH radical scavenging activity, ${\alpha}$-glucosidase and tyrosinase inhibitory assay (used ${\text\tiny{L}}$-tyrosine as substrate), the 80% ethanol extract exhibited a higher value than other extracts. The 60% ethanol extract showed prominent activities in the tyrosinase inhibitory (used ${\text\tiny{L}}$-dopa as substrate), ADH and ALDH activity assay. In the minimum inhibitory concentration (MIC) assay, 60% and 80% ethanol extracts inhibited the bacterial growth almost similarly. Moreover, the gram-positive bacteria was more restrained than the gram-negative bacteria. The resultsrevealed that the distilled water and 80% ethanol extract showed a relatively higher antioxidant activity compared to other extracts. The 60 ~ 80% ethanol extracts demonstrated potential tyrosinase, ${\alpha}$-glucosidase inhibitory, antimicrobial, ADH and ALDH activities. Therefore, the C. grandis is suggested to be considered as a functional material for various proposes.

Antilisterial activity of fresh cheese fermented by Lactobacillus paracasei BK57 (Lactobacillus paracasei BK57 균주로 발효시킨 프레쉬 치즈의 항리스테리아 활성)

  • Lim, Eun-Seo;Lee, Eun-Woo
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.407-418
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    • 2015
  • This study is focused on establishing the optimal conditions to enhance the production of antilisterial substances by Lactobacillus paracasei BK57 isolated from Baikkimchi. In addition, the growth and in situ lactic acid and bacteriocin production of this strain were investigated during the manufacture of fresh cheese. And then the efficacy of using Lactobacillus starter as a protective culture to improve the safety of fresh cheese against Listeria monocytogenes KCTC 3569 was estimated. Maximum growth rate and activity of antibacterial substances were obtained in Lactobacilli MRS broth at $37^{\circ}C$ with controlled pH 6.0 after 30 h of incubation under aerobic condition. However, the growth rate and antimicrobial activity of bacteriocin produced in whole milk supplemented with yeast extract (2.0%) as a substrate were lower than those obtained in MRS broth. Live cells and cell-free culture supernatant of BK57 strain were effective in the suppression of L. monocytogenes in milk, whereas the inhibitory of the bacteriocin obtained from BK57 strain was higher in BHI broth than in milk. During storage at $4^{\circ}C$ and $15^{\circ}C$ for 6 days, no significant difference was found in the cell viability and antimicrobial activity of BK 57 strain in fresh cheese. In samples held at two temperatures, there was at least a 15% reduction in the numbers of the pathogen in fresh cheese artificially contaminated with approximately $10^5CFU/ml$ of L. monocytogenes within 6 days. Our results demonstrated the usefulness of L. paracasei BK57 having antilisterial activity as a biopreservative in the cheese making process.

In Vitro Antagonistic Activity Evaluation of Lactic Acid Bacteria (LAB) Combined with Cellulase Enzyme Against Campylobacter jejuni Growth in Co-Culture

  • Dubois-Dauphin, Robin;Sabrina, Vandeplas;Isabelle, Didderen;Christopher, Marcq;Andre, Thewis;Philippe, Thonart
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.62-70
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    • 2011
  • The antibacterial effects of nine lactic acid bacteria (LAB) against Campylobacter jejuni were investigated by using agar gel diffusion and co-culture assays. Some differences were recorded between the inhibition effects measured with these two methods. Only two LAB, Lb. pentosus CWBI B78 and E. faecium THT, exhibited a clear anti- Campylobacter activity in co-culture assay with dehydrated poultry excreta mixed with ground straw (DPE/GS) as the only growth substrate source. It was observed that the supplementation of such medium with a cellulase A complex (Beldem S.A.) enhanced the antimicrobial effect of both LAB strains. The co-culture medium acidification and the C. jejuni were positively correlated with the cellulase A concentration. The antibacterial effect was characterized by the lactic acid production from the homofermentative E. faecium THT and the lactic and acetic acids production from the heterofermentative Lb. pentosus CWBI B78. The antagonistic properties of LAB strains and enzyme combination could be used in strategies aiming at the reduction of Campylobacter prevalence in the poultry production chain and consequently the risk of human infection.