• Title/Summary/Keyword: bacterial enzymes

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Studies on Microbial and Enzymatic Actions during the Ripening Process of Salted Alaska Pollack Tripe (창난 젓갈의 숙성 과정 중 미생물 및 자기소화효소 작용에 관한 연구)

  • Chae, Soo-Kyu
    • The Korean Journal of Food And Nutrition
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    • v.24 no.3
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    • pp.340-349
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    • 2011
  • This study examined the roles of autolytic enzymes and microorganisms in the ripening process of salted Alaska pollack tripe made with various concentrations of salt i.e, 7.5% and 20% by weight. Salted Alaska pollack tripe treated with antibiotic agents for the inhibition of microbial growth and a control were prepared experimentally, and changes in chemical composition and viable cell counts were investigated, individually, during the ripening process. Just after the preparation of the low salt Alaska pollack tripe made with 7.5% salt, viable bacterial cells occurred at a level of $10^5$ CFU/g. In the control, bacterial counts increased rapidly to $10^7$ CFU/g by the 14th day of ripening. However, in the sample treated with antibiotic agents, counts were decreased to a level of $10^4$ CFU/g by the 3rd day of ripening and increased gradually to $10^6$ CFU/g by the 5th day of ripening, and then the same value was maintained there-after. Just after the preparation of the high salt Alaska pollack tripe made with 20% salt, viable bacterial cells occurred at a level of $10^3$ CFU/g. In both the samples treated with antibiotic agents and the control, bacterial counts decreased rapidly to $10^0$ CFU/g by the 45th day of ripening and increased gradually there-after. The content of amino type nitrogen was 76.3 mg% just after the preparation of the low salt Alaska pollack tripe made with 7.5% salt. Amino type nitrogen content was increased to 283.5 mg% by the 5th day of proper ripening in the control, but it was increased to 208.0 mg% in the sample treated with antibiotic agents. The difference in amino type nitrogen content was 75.5 mg/100 g. The content of amino type nitrogen was 57.2 mg% just after the preparation of the high salt Alaska pollack tripe made with 20% salt. Amino type nitrogen content was increased to 198.3 mg by the 60th day of proper ripening in the control, but it was increased to 162.0 mg% in the sample treated with the antibiotic agents. The difference in amino type nitrogen content was 36.3 mg/100 g. The contents of VBN and TMA-N were 102.1 mg% and 20.5 mg%, respectively, at the 7th day of ripening in the low salt Alaska pollack tripe made with 7.5% salt. The content of VBN was 60.0 mg% and TMA-N was not detected at the 21st day of ripening in the sample treated with antibiotic agents. The control sample was spoiled by the 7th day of ripening but the sample treated with antibiotic agents was not spoiled by the 21st day of ripening. On the other hand, VBN content was 37.2 mg% and TMA-N was not detected at the 90th day of ripening in the high salt Alaska pollack tripe made with 20% salt, and the control sample was not spoiled.

Activation of RAW 264.7 Macrophage by Digested Bacterial Cell of Pig-derived Lactobacillus Strains (돼지에서 분리된 Lactobacillus Strains의 균체분해산물에 의한 RAW 264.7 Macrophage 활성화)

  • Kim, D.W.;Cho, S.B.;Jeong, H.Y.;Moon, H.G.;Lee, H.J.;HwangBo, J.;Chung, W.T.;Choi, C.W.;Chung, I.B.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.947-954
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    • 2005
  • This study was conducted to investigate the effects of hydrolyzed Lactobacillus supplementation with digestive enzymes treatment on the macrophage activation, the induction of nitric oxide(NO), interleukin (IL)-6 and tumor necrosis factor(TNF)-$\alpha$. The RAW 264.7 murine macrophage was exposed to porcine Lactobacillus strains which were digested with both pepsin and pancreatin. The production of NO, TNF-$\alpha$ and IL-6 in the macrophage were strain and dose-dependent, respectively. The induction of NO and cytokines were higher in both 3149 and 3146 strains compared with other Lactobacillus strains. Overall, the level of NO was observed at the lower range between 10 and 150 μg hydrolysates per ml, whereas IL-6 and TNF-$\alpha$ were observed at relatively higher concentration between 50 and 300 μg hydrolysates per ml. Lactobacillus strains which produced a high level of NO also showed a high induction of TNF-$\alpha$ and IL-6. Therefore, the present results suggest that hydrolysates of Lactobacillus strains are related to induction of several macrophage mediators, and then it could be beneficially used to modulate gastrointestinal immune function of the host. Also, the methodogly employed in this study might be useful to investigate the effects of lactic acid bacteria on gastrointestinal immunity.

Diversity and Physiological Characteristics of Culturable Bacteria from Marine Sediments of Ross Sea, Antarctica (남극 로스해 퇴적물로부터 분리된 세균의 다양성 및 생리학적 특성)

  • Lee, Yung Mi;Jung, You-Jung;Hong, Soon Gyu;Kim, Ji Hee;Lee, Hong Kum
    • Korean Journal of Microbiology
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    • v.50 no.2
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    • pp.119-127
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    • 2014
  • The affiliations and physiological characteristics of culturable bacteria isolated from the sediments of Ross Sea, Antarctica were investigated. Sixty-three isolates obtained by cultivation were grouped into 21 phylotypes affiliated with the phyla Actinobacteria and Bacteroidetes and with the classes Alphaproteobacteria and Gammaproteobacteria by phylogenetic analysis of 16S rRNA gene sequences. Based on phylogenetic analysis (<98.65% sequence similarity), approximately 49% of total isolates represented potentially novel species or genus. Among them, extracellular protease, lipase, and exopolysaccharide activities at $10^{\circ}C$ or $20^{\circ}C$ were detected in approximately 46%, 25%, and 32% of the strains, respectively. Forty-three isolates produced at least one type of extracellular material and 21 of them produced at least two extracellular protease, lipase, and/or exopolysaccharides. Our findings indicate that culturable bacterial diversity present within the marine sediments of Ross Sea, Antarctica may contribute to the hydrolysis of the major organic constituents which is closely related with carbon and nitrogen cycling in this environment.

Production and Characterization of α-Galactosidases from Two Bacillus licheniformis Isolates (Bacillus licheniformis 분리균 2종의 α-Galactosidase 생산성과 효소특성)

  • Jin, Hyun Kyung;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.195-203
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    • 2015
  • Two bacterial strains, Bacillus licheniformis YB-1413 and YB-1414, producing extracellular α-galactosidase, were obtained from homemade Doenjang. On the basis of their biochemical properties, 16S rRNA sequences and random amplified polymorphic DNA patterns by polymerase chain reaction, they were found to be somewhat different from one another. α-Galactosidase productivities of the two isolates were increased by wheat bran, but drastically decreased by melibiose, raffinose and sucrose which were used as carbon sources. The enzyme productivities were increased by yeast extract as a nitrogen source with maximum levels of 1.87 U/ml for YB-1413 and 1.69 U/ml for YB-1414, respectively. The enzymes of both isolates exhibited maximum activity for hydrolysis of para-nitrophenyl-α-D-galactopyranoside (pNP-αGal) under reaction conditions of pH 6.0 and 45℃. Their hydrolyzing activities for pNP-αGal were drastically decreased by the addition of low concentrations of ribose and galactose. They were capable of hydrolyzing completely α-1,6 linked galactosyl residue in melibiose, raffinose and stachyose, which are known to be anti-nutritional factors in products of soybean and legume. In relation to the latter, the isolates YB-1413 and YB-1414 have potential applicability in improving soybean-fermented foods and the nutritional value of soybean feed.

Characterization of Antimicrobial Substance Produced by Lactobacillus sp. HN 235 Isolated from Pig Intestine (돼지 장관으로부터 분리한 Lactobacillus sp. HN 235 균주가 생산하는 항균물질의 특성)

  • Shin, Myeong-Su;Han, Sun-Kyung;Choi, Ji-Hyun;Ji, Ae-Ran;Kim, Kyeong-Su;Lee, Wan-Kyu
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.125-132
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    • 2009
  • In order to develop probiotics which may be a viable alternative of antibiotic use in pig industry, five bacterial strains (Lactobacillus sp. HN 52, 92, 98, 235 and AP 116) possessing antimicrobial properties were selected from 500 strains isolates of pig intestines. The bacteriocin produced by Lactobacillus sp. HN 235 displayed a relative broad spectrum of inhibitory activity against all Enterococcus strains, Pseudomonas aeruginosa, Listeria monocytogenes and Clostridium perfringens using the spot-on-lawn method. The production of antimicrobial substance started in the middle of the exponential growth phase, reached maximum levels (6,400 AU/mL) in the stationary phase, and then declined. Bacteriocin activity remained unchanged after 30 min of heat treatment at $95^{\circ}C$ and stable from pH 2.0 to 10 for 1 h, or exposure to organic solvents; however, it diminished after treatment with proteolytic enzymes. The molecular weight of the bacteriocin was about 5 kDa according to a tricine SDS-PAGE analysis.

Antibacterial Effect of Antibacterial Substance Produced by Lactobacillus amylovorus IMC-1 against Food Spoilage Bacteria (Lactobacillus amylovorus IMC-1에 의해서 생산되는 항균성 물질의 식품 오염세균에 대한 항균 효과)

  • Mok, Jong-Soo;Kim, Poong-Ho;Yu, Hyen-Duk;Kim, Ji-Hoe;Lee, Hee-Jung;Kim, Young-Mog
    • Journal of Food Hygiene and Safety
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    • v.14 no.4
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    • pp.346-351
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    • 1999
  • To develop a lactic starter to produce antimicrobial substance for inhibiting the growth of a variety of foodborne spoilage bacteria in fermented foods, we investigated the anti-bacterial effect of the antibacterial substance, produced by Lactobacillus amylovorus IMC-1, against foodborne spoilage strains, and its sensitivity on the treatment of proteolytic enzymes. L. amylovorus IMC-1, which was isolated from a traditional cheese in Inner Mongolia, produced a maximum amount of antibacterial substance in the skim milk medium after 72 h incubation at 37$^{\circ}C$, and further incubation resulted in the same activity. The substance obtained from gel filtration inhibited all strains used such as Bacillus subtilis IFO 3025, Staphylococcus aureus IAM 1011, Listeria monocytogenes VTU 206, Escherichia coli RB, and Pseudomonas fragi IFO 3458 at the concentration of 20 units/ml. This substance was found to show bactericidal action against B. subtilis, E. coli, and Ps. fragi, and bacteriostatic activity against both Staph. aureus and L. monocytogenes. The bactericidal action was due to cellular Iysis. The substance is not organic acid, hydrogen peroxide and proteinaceous compound.

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Anti-inflammatory and antimicrobial effects of Jeju rosemary essential oil against skin flora (제주산 로즈마리 에센셜 오일의 항염 및 피부 상재균에 대한 항균 활성)

  • Kim, So-Hee;Yi, Mi-Ran;Kim, Chang-Soo;Kim, Jung-Mi;Bu, Hee-Jung
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.3
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    • pp.744-756
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    • 2018
  • The purpose of this study was to investigate the anti-inflammatory effect and the antimicrobial activity to skin flora of essential oil from rosemary that naturally grown in Jeju. rosemary essential oil was extracted by water distillation essential oil extraction method. In order to confirm the anti-inflammatory activity of rosemary essential oil, it was confirmed that the production of NO and $PGE_2$ induced by LPS in RAW 264.7 cells was inhibited in a concentration-dependent manner. Western blot analysis showed that the expression of iNOS and COX-2, which are biosynthetic enzymes, decreased in a concentration-dependent manner. In addition, production of $TNF-{\alpha}$ and IL-6 the pro-inflammatory cytokines were inhibited. Antimicrobial activities of three S. epidermidis and three P. acnes strains including two antibiotic resistant strains were observed in paper disc method and MIC and MBC tests showed inhibition of bacterial growth and death. From the results of the experiment, we confirmed that rosemary essential oil has the anti-inflammatory and antimicrobial efficacy and it could be used as a cosmetic and skin care material in the future.

Identification and Characterization of an Agarase- and Xylanse-producing Catenovulum jejuensis A28-5 from Coastal Seawater of Jeju Island, Korea (제주 연안해수로부터 한천 분해 효소 및 자일란 분해 효소를 생산하는 Catenovulum jejuensis A28-5의 동정 및 특성 규명)

  • Kim, Da Som;Jeong, Ga Ram;Bae, Chang Hwan;Yeo, Joo-Hong;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.168-177
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    • 2017
  • Strain A28-5, which can degrade xylan and agar in solid medium, was isolated from a coastal seawater sample collected from Jeju Island, South Korea. This strain was found to be a gram-negative, $Na^+$-requiring bacterial strain with a polar flagellum for motility. Additionally, the strain was tolerant to antibiotics such as ampicillin and thiostrepton. The G+C content of the genome was 43.96% and menaquinone-7 was found to be the predominant quinone. Major fatty acids constituting the cell wall of the strain were $C_{16:1}$ ${\omega}7c/iso-C_{15:0}$ 2-OH (23.32%), $C_{16:0}$ (21.83%), and $C_{18:1}$ ${\omega}7c$ (17.98%). The 16S rRNA gene sequence of the strain showed the highest similarity (98.94%) to that of Catenovulum agarivorans YM01, which was demonstrated by constructing a neighbor-joining phylogenetic tree. A28-5 was identified as a novel species of the genus Catenovulum via DNA-DNA hybridization with Catenovulum agarivorans YM01, and thus was named as Catenovulum jejuensis A28-5. The formation of tetramers and hexamers of xylooligosaccharides and (neo)agarooligosaccharides, respectively, were confirmed by thin-layer chromatography analysis using an enzyme reaction solution containing xylan or agarose with two crude enzymes prepared from the liquid culture of the strain.

Interactions between Biosynthetic Pathway and Productivity of IAA in Some Rhizobacteria (근권에서 분리한 세균의 IAA 생합성 경로와 IAA 생성능과의 관계)

  • Kim, Woon-Jin;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.48 no.1
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    • pp.1-7
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    • 2012
  • This study explores the interaction between the production of indole-3-acetic acid (IAA), a typical phytohormone auxin and the role of IAA biosynthetic pathways in each IAA producing rhizobacterial strain. The bacterial strains were isolated from rhizosphere of wild plants and identified as Acinetobacter guillouiae SW5, Bacillus thuringiensis SW17, Rhodococcus equi SW9, and Lysinibacillus fusiformis SW13. A. guillouiae SW5 exhibited the highest production of IAA using tryptophan-dependent pathways among the 4 strains. When indole-3-acetamide (IAM) was added, Rhodococcus equi SW9 showed the highest IAA production of $3824{\mu}g/mg$ protein using amidase activity. A. guillouiae SW5 also showed the highest production of IAA using two pathways with indole-3-acetonitrile (IAN), and its nitrile hydratase activity might be higher than nitrilase. B. thuringiensis SW17 showed the lowest IAA production, and most of IAA might be produced by the amidase activity, although the nitrilase activity was the highest among 4 strains. The roles of nitrile converting enzymes were relatively similar in IAA synthesis by Lysinibacillus fusiformis SW13. Tryptophan-independent pathway of IAA production was utilized by only A. guillouiae SW5.

Anti-inflammatory Effects of Ethanolic Extracts from Codium fragile on LPS-Stimulated RAW 264.7 Macrophages via Nuclear Factor kappaB Inactivation

  • Yoon, Ho-Dong;Jeong, Eun-Ji;Choi, Ji-Woong;Lee, Min-Sup;Park, Myoung-Ae;Yoon, Na-Young;Kim, Yeon-Kye;Cho, Deuk-Moon;Kim, Jae-Il;Kim, Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • v.14 no.4
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    • pp.267-274
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    • 2011
  • Bacterial lipopolysaccharide (LPS) induces expression of pro-inflammatory cytokines and enzymes producing nitric oxide (NO) and prostaglandins (PGs) in immune cells. This process is mediated by the activation of nuclear factor kappaB (NF-${\kappa}B$). In this study, we investigated the anti-inflammatory characteristics of Codium fragile ethanolic extract (CFE) mediated by the regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) using LPS-stimulated murine macrophage RAW 264.7 cells. CFE significantly inhibited LPS-induced NO and $PGE_2$ production in a dose-dependent manner and suppressed the expression of iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells with no cytotoxicity. Pro-inflammatory cytokines, such as interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor-${\alpha}$, were significantly reduced by treatment of CFE in LPS-stimulated RAW 264.7 cells. CFE inhibited the promoter activity of (NF)-${\kappa}B$ in LPS-stimulated macrophages. Treatment with CFE suppressed translocation of the NF-${\kappa}B$ p65 subunit by preventing proteolytic degradation of inhibitor of ${\kappa}B-{\alpha}$. These results indicate that the CFE-mediated inhibition of NO and $PGE_2$ production in LPS-stimulated RAW 264.7 cells is mediated through the NF-${\kappa}B$-dependent transcriptional downregulation of iNOS and COX-2, suggesting the potential of CFE as a nutraceutical with anti-inflammatory activity.