• Title/Summary/Keyword: Bacterial mixture

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Determination of Optimized Growth Medium and Cryoprotective Additives to Enhance the Growth and Survival of Lactobacillus salivarius

  • Yeo, Soyoung;Shin, Hee Sung;Lee, Hye Won;Hong, Doseon;Park, Hyunjoon;Holzapfel, Wilhelm;Kim, Eun Bae;Huh, Chul Sung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.718-731
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    • 2018
  • The beneficial effects of lactic acid bacteria (LAB) have been intensively investigated in recent decades with special focus on modulation of the host intestinal microbiota. Numerous discoveries of effective probiotics are driven by a significantly increasing demand for dietary supplements. Consequently, technological advances in the large-scale production and lyophilization are needed by probiotic-related industries for producing probiotic LAB for commercial use. Our study had a dual objective, to determine the optimum growth medium composition and to investigate appropriate cryoprotective additives (CPAs) for Lactobacillus salivarius, and compare its responses with other Lactobacillus species. The one-factor-at-a-time method and central composite design were applied to determine the optimal medium composition for L. salivarius cultivation. The following composition of the medium was established (per liter): 21.64 g maltose, 85 g yeast extract, 1.21 ml Tween 80, 6 g sodium acetate, $0.2g\;MgSO_4{\cdot}7H_2O$, $0.02g\;MnSO_4{\cdot}H_2O$, $1g\;K_2HPO_4$, $1.5g\;KH_2PO_4$, $0.01g\;FeSO_4{\cdot}7H_2O$, and 1 g sodium citrate. A cryoprotective additive combination comprising 10% (w/v) skim milk and 10% (w/v) sucrose supplemented with 2.5% (w/v) sodium glutamate was selected for L. salivarius, and its effectiveness was confirmed using culture-independent methods in the freeze-dried cells of the Lactobacillus strains. In conclusion, the optimized medium enhanced the species-specific cultivation of L. salivarius. On the other hand, the cryoprotective effects of the selected CPA mixture may also be dependent on the bacterial strain. This study highlights the necessity for precise and advanced processing techniques for large-scale production of probiotics in the food and feed industries.

Effect of Exogenous Fibrolytic Enzyme Application on the Microbial Attachment and Digestion of Barley Straw In vitro

  • Wang, Y.;Ramirez-Bribiesca, J.E.;Yanke, L.J.;Tsang, A.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.66-74
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    • 2012
  • The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

Evaluation of Medicinal Activity on Isolated Inhibitory Compounds against Helicobacter pylori from Cheongmoknosang Mulberry Leaves (청목노상 뽕잎에서 분리한 Helicobacter pylori 저해물질에 대한 약효평가)

  • Kim, Byong-O;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.54 no.4
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    • pp.265-269
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    • 2011
  • Inhibitory effect of useful components from Cheongmoknosang mulberry leaves extracts against Helicobacter pylori were investigated to develope them to a health functional food. It was confirmed that H. pylori bacterial infection occurred after 6 weeks over in C57BL/6 mouse which was caused the infection, and the average number of pathogens was $8{\times}10^5$ CFU/mL. Effects of the prevention and cure against H. pylori were tested by the mouth administration with Cheongmoknosang mulberry leaves extracts include the active ingredient, and number of H. pylori colony in stomach of drug groups were decreased more than control group. The result of testing immuno-glogulin isotype from the separated serum from a mouse, IgG1, IgA produced more in administered group, and it is expected to inhibit the H. pylori infection because of increasing antibody production in the mixture. These results suggest that caffeic acid, rosemarinic acid and chlorogenic acid in Cheongmoknosang mulberry leaves extracts are very effective to prevent or cure against H. pylori infection. So the ratio of infection is increasing and it is regarded to be able to prevent and cure the disease like gastritis, gastric ulcer and gastric cancer which are caused by H. pylori.

Cometabolism of $\omega$-Phenylalkanoic Acids with Butyric Acid for Efficient Production of Aromatic Polyesters in Pseudomonas putida BM01

  • Song, Jae-Jun;Choi, Mun-Hwan;Yoon, Sung-Chul;Huh, Nam-Eung
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.435-442
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    • 2001
  • Poly(3-hydroxy-5-phenylvalerate) [P(3HPV)] was efficiently accumulated from 5-phenylvalerate (5PV) in Pseudomonas putida BM01 in a mineral salts medium containing butyric acid (BA) as the cosubstrate. A nove aromatic copolyester, poly(5 mol% 3-hydroxy-4-phenylbutyrate-co- 95 mol% 3-hydroxy-6-phenylhexanoate) [P(3HPB-co-3HPC)] was also synthesized from 6-phenylhexanoate (6PC) plus Ba. The two aromatic polymers, P(3HPV) and P(3HPB-co-3HPC), were found to be amorphous and showed different glass-transition temperatures at $15^{\circ}C$ and $10^{\circ}C$, respectively. When the bacterium was grown ina medium containing 20 mM 5PV as the sole carbon source for 140 h, 0.4 g/l of dry cells was obtained in a flask cultivation and 20 wt% of P(3HPV) homopolymer was accumulated in the cells. However, when it was grown with a mixture of 2 mM 5PV and 50 mM BA for 40 h, the yield of dry biomass was increased up to 2.5 g/l and the content of P(3HPV) in the dry cells was optimally 56 wt%. This efficient production of P(3HPV) homopolymer from the mixed substrate was feasible because BA only supported cell growth and did not induce any aliphatic PHA accumulation. The metabolites released into the PHA synthesis medium were analyzed using GC or GC/MS. Two $\beta$-oxidation derivatives, 3-phenylpropionic acid and trans-cinnamic acid, were found in the 5V-grown cell medium and these comprised 55-88 mol% of the 5PV consumed. In the 6PC-grown medium containing Ba, seven ${\beta}$-oxidation and related intermediates were found, which included phenylacetic acid, 4-phenylbutyric acid, cis-4-phenyl-2-butenoic acid, trans-4-phenyl-3-butenoic acid, trans-4-phenyl-2-butenoic acid, 3-hydroxy-4-phenylbutyric acid, and 3-hydroxy-6-phenylhexanoic acid. Accordingly, based on the metabolite analysis, PHA synthesis pathways from the two aromatic carbon sources are suggested.

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Development of Antimicrobial Plant Extracts and its Application to Cosmetics (항균활성을 갖는 천연물 개발과 화장품 응용에 관한 연구)

  • Lee, Ji-Young;Lee, Jung-No;Lee, Ghang-Tai;Lee, Kun-Kook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.2
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    • pp.171-179
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    • 2012
  • This study is focused on finding new natural materials that have antimicrobial activity. We found that 3 plants extracts, Magnolia sieboldii K. Koch (M. sieboldii K. Koch), Rhus chinensis (R. chinensis) and Metasequioa glyptostroboides (M. glyptostroboides), have antimicrobial activities by disk diffusion method. We determined the Minimum Inhibitory Concentation (MIC) of each and found that 0.3 ~ 0.35 % of M. glyptostroboides essential oil and 0.35 ~ 0.4 % of M. sieboldii K. Koch extracts inhibited fungal growth and 0.45 ~ 0.5 % of R. chinensis extracts inhibited bacterial growth. We isolated compounds from extracts and verified what have antimicrobial activity. As a result we found that caryophyllene oxide and caryophyllene isolated from M. glyptostroboides, costunolide and dehydrocostus lactone from M. sieboldii K. Koch and ethyl gallate, ethyl-3-gallate from R. chinensis have antimicrobial activities. In accordance with antimicrobial activity, O/W cosmetic emulsion containing mixture of 3 plants extracts showed preservative efficacy against both bacteria and fungi. Based on the above data we suggest that extracts from M. siebodii K. Koch, R. chinensis, and M. glyptostroboides, replace chemical synthetic preservatives and be applied as a natural preservative.

Biological Control of Root-Knot Nematodes by Organic Acid-Producing Lactobacillus brevis WiKim0069 Isolated from Kimchi

  • Seo, Hye Jeong;Park, Ae Ran;Kim, Seulbi;Yeon, Jehyeong;Yu, Nan Hee;Ha, Sanghyun;Chang, Ji Yoon;Park, Hae Woong;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.662-673
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    • 2019
  • Root-knot nematodes (RKNs) are among the most destructive plant-parasites worldwide, and RKN control has been attempted mainly using chemical nematicides. However, these chemical nematicides have negative effects on humans and the environment, thus necessitating the search for eco-friendly alternative RKN control methods. Here, we screened nematicidal lactic acid bacteria (LAB) isolated from kimchi and evaluated their efficacy as biocontrol agents against RKNs. Of 237 bacterial strains, Lactobacillus brevis WiKim0069 showed the strongest nematicidal activity against the second-stage juveniles (J2) of Meloidogyne incognita, M. arenaria, and M. hapla and inhibited the egg hatch of M. incognita. The culture filtrate of WiKim0069 had a pH of 4.2 and contained acetic acid (11,190 ㎍/ml), lactic acid (7,790 ㎍/ml), malic acid (470 ㎍/ml), and succinic acid (660 ㎍/ml). An artificial mixture of the four organic acids produced by WiKim0069 also induced 98% M. incognita J2 mortality at a concentration of 1.25%, indicating that its nematicidal activity was derived mainly from the four organic acids. Application of WiKim0069 culture filtrate suppressed the formation of galls and egg masses on tomato roots by M. incognita in a dose-dependent manner in a pot experiment. The fermentation broth of WiKim0069 also reduced gall formation on melon under field conditions, with a higher efficacy (62.8%) than that of fosthiazate (32.8%). This study is the first report to identify the effectiveness of kimchi LAB against RKNs and to demonstrate that the organic acids produced by LAB can be used for the RKN management.

Effect of Dietary Supplementation of Sodium Salt of Isobutyric Acid on Ruminal Fermentation and Nutrient Utilization in a Wheat Straw Based Low Protein Diet Fed to Crossbred Cattle

  • Misra, A.K.;Thakur, S.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.4
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    • pp.479-484
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    • 2001
  • The effect of dietary supplementation of sodium salt of isobutyric acid in low protein (10% CP) wheat straw based diet on nutrient utilization and rumen fermentation was studied in ruminally fistulated male crossbred cattle. The study included a 7 day metabolism and a 3 day rumen fermentation trials. The cattle were distributed into two equal groups of 4 each. The animals of control group were fed a basal diet consisting of wheat straw, concentrate mixture and green maize fodder in 40:40:20 proportion whereas branched chain volatile fatty acid (BCFA) supplemented group received a basal diet + isobutyric acid at 0.75 percent of basal diet. The duration of study was 36 days. The feed intake between experimental groups did not differ significantly and the average total DMI (% BW) was 2.01 and $2.28kg\;day^{-1}$ in control and BCFA supplemented diets. The dietary supplementation of BCFA improved (p<0.05) the DM, OM, NDF and cellulose digestibility by 4.46, 6.63, 10.57 and 11.31 per cent over those fed control diet. The total N retention on BCFA supplementation was improved (p<0.01) due to decreased (p<0.05) urinary N excretion. The concentrations of ruminal total N was 37.07 and $34.77mg\;100ml^{-1}$ in control and BCFA fed groups, respectively. Dietary supplementation BCFA significantly (p<0.01) reduced the ruminal ammonia N concentration as compared to control and the mean values ($mg\;100ml^{-1}$) were 13.18 and 9.42 in control and BCFA fed groups. The total VFA concentration was higher (p<0.01) in BCFA supplemented group (101.14 mM) than the control (93.05 mM). Among the VFAs, the molar proportion of acetate was higher (p<0.01) in BCFA supplemented group (71.07 mM) as compared to control (64.98 mM). However, the concentration of propionate and butyrate remained unchanged. Amino acids composition of bacterial hydrolysates was similar in both the groups. Ruminal outflow rate of liquid digesta was higher (p<0.01) in BCFA fed group ($67.56l\;day^{-1}$) than control ($52.73l\;day^{-1}$). It is concluded that the dietary supplementation of Na-salt of isobutyric acid in low protein diet improved the nutrient utilization and ruminal fermentation characteristics.

ANALYSIS OF PARA-CHLOROANILINE AFTER CHEMICAL INTERACTION BETWEEN ALEXIDINE AND SODIUM HYPOCHLORITE USING MASS SPECTROMETRY (알렉시딘과 차아염소산나트륨의 화학적 상호반응 후 mass spectrometry를 이용한 파라클로로아닐린의 검출 분석)

  • Kim, Hyeon-Sik;Han, Seung-Hyun;Oh, So-Ram;Lim, Sang-Min;Gu, Yu;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.35 no.4
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    • pp.295-301
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    • 2010
  • Recent studies demonstrated that the combination of chlorhexidine (CHX) and sodium hypochlorite (NaOCl) resulted in the formation of a precipitate, para-chloroaniline (PCA). Alexdidine (ALX) is a kind of biguanides like CHX, and has stronger detoxification effect against the bacterial virulence factors such as lipoteichoic acid and lipopolysacchardide compared with CHX. The purpose of this study was to determine whether PCA was formed after chemical interaction between ALX and NaOCl using mass spectrometry. Mass spectrometry was performed for the mixture of five different concentrations of ALX (1, 0.5, 0.25, 0.125, 0.0625%) and 4% NaOCl. Results showed that the peak of PCA was not detected in mixed solutions of ALX and NaOCl in mass spectrometry analysis. The color of mixed solution of ALX and NaOCl after chemical interaction was light yellow to white, but there wasn't any precipitate found.

Studies on the EPA(eicosapentaenioc acid) production by marine bacteria (해양세균(海洋細菌)을 이용(利用)한 EPA(eicosapentaenoic acid) 생성(生成)에 관한 연구(硏究))

  • Joh, Yong-Goe;Kim, Seong-Jin;Kim, Ji-Soo;Lee, Min-Kyeng
    • Journal of the Korean Applied Science and Technology
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    • v.12 no.2
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    • pp.121-130
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    • 1995
  • This project has been worked out for isolation of EPA-producing bacteria from marine source of sea water, sea sediment and intestinal contents eviscerated from some red-muscle fish such as mackerel, horse-mackerel and spike fish. The samples were precultured on the media of PPES-II glucose broth and then pure-cultured on Nutrient agar and P-Y-M glucose. Lipids extracted from those bacterial mass collected by centrifugation were analysed in terms of lipid class and fatty acid composition. The results are resumed as follows : 1. 112 strains from sea water and 76 strains from sea sediment were tested for their EPA producing capability, but both strains of (SA-67 and SA-91) from the former and four strains(SS-35, 37, 51 and 71) from the latter have been proved to produce EPA above the level of 2% of total fatty acids. The strains such as GS-11, 29, 31, HM-9, 29, B-18, 33, 107, YL-129, 156, 203, 77, 104 and 256 which were isolated from fish intestinal contents, have also produced EPA at higher level than 2% of total fatty acids. 2. Contents of total lipids extracted from the cultures of these strains grown at $25^{\circ}C$, range from 2.8% to 6.9% (on dry weight %), and they are mainly composed of polar lipids($40.9{\sim}52.9%$) such as phosphatidyl glycerol($^{+}cardiolipin$)(?) and phosphatidyl ethanolamine ($33.8{\sim}40.0%$), with smaller amount of free fatty acid ($11.2{\sim}20.2%$). 3. EPA was isolated from a mixture of fatty acid methyl esters obtained from the lipid of each strain by HPLC in silver-ion mode and was identified by GC-Mass spectrometry. 4. The strains of SW-91, GS-11, GS-29, HM-9, B-18 and YL-203 grown at $25^{\circ}C$ have a level of 5% EPA in their total fatty acids, and the GS-11 and HM-9 strains show a tendency of increase in the EPA level with an increase of growth temperature.

The Growth Promotion Effect of Useful Enterobacteria Bifidobacterium aolescentis KCTC 3216 by Combination of Natural Products Bearing Antioxioative Capacity (장내 유용세근 Bifidobacterium adolescentis KCTC 3216의 성장을 촉진시키는 항산화 천연산물의 조합구성)

  • 김종덕;김민용;안창범;서효진;김봉조;서재관;김점순;공재열
    • KSBB Journal
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    • v.17 no.4
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    • pp.388-395
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    • 2002
  • The growth of enterobacteria, Bifidobacteriurn adolescentis KCTC 3216 was promoted by natural products bearing antioxidative capacity and combined two, three and four kinds of them. B. adolescentis was showed a good growth by Sophorae flos, Phellodendri cofex, Mori cortex radicis, Aurantii nobilis pericarpium, Angelicae gigantis radix, alone, and two mixed combinations were composed of Paeonia japonica and Theae foiium, Epimedii herba and Angelicae gigantis radix, Paeonia japonica and epimedii herba, Atractylodis rhizoma alba and angelicae gigantis radix, and three mixed combinations were oraganized with Theae folium, Paeonia japonica and epimedii herba, Theae folium, Beiamcanda chinensis and Paeonia japonica, Theae foiium, Astragaii radix and Mori cortex radicis, and four mixed combinations were formed with Seiamcanda chinensis, Angelicae gigantis radix, Epimedii herba, Theae folium, and Angeiicae gigantis radix, Epimedii herba, Paeonia japonica, Theae folium, and Epimedii herba, Paeonia japonica, Sophorae flos, Theae folium. The best four mixed combination for the growth of B. adolescentis was mixture of Epimedii herba, Paeonia iaponica, Sophorae flow and Theae foiium, which promoted 2.6 times than that of control, and its antioxidative capacity was also 5.6 times higher, and the ratio of elimination of hydroxyl radical was more than 80% in each dilution rate. As these combinations of natural products will activate some parts of body, they may be applied to pharmaceuitcal applications, functional foods, antiaging tea, also expected to promote useful entero bacterial growth for fermentative beverage bearing multifunction.