• Title/Summary/Keyword: $^1H-MRS$

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Characterization of Cholesterol Lowering Lactic Acid Bacteria Isolated from Palm Wine and Maize Beer and Assessment of Their Use in the Production of Probiotic Papaya Juice

  • Bertrand Tatsinkou Fossi;Dickson Ebwelle Ekabe;Liliane Laure Toukam Tatsinkou;Rene Bilingwe Ayiseh;Frederic Tavea;Pierre Michel Jazet
    • Microbiology and Biotechnology Letters
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    • v.51 no.2
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    • pp.191-202
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    • 2023
  • Elevated serum cholesterol is a main risk factor for heart disorders. Most probiotic products administered to lower cholesterol are dairy products which are not suitable for lactose-intolerant individuals. In this study, we assessed the cholesterol-lowering efficacy of LAB isolated from traditionally fermented drinks in diet-induced rats and determine their efficacy in the production of non-dairy, probiotic formulations using papaya juice. LAB were isolated from palm wine and corn beer on MRS agar using a pour-plate technique. Identification was carried out using 16S rRNA gene sequencing. A hypercholesterolemia model in which diet-induced Wistar albino rats were assigned into four groups was established. Oral gavage was carried out for 30 days. On the 31st day, the rats were dissected and the serum lipid profile was analyzed using biochemical kits. A 106 cfu/ml of a 24-h-old culture of selected lactobacilli was used to inoculate papaya juice and incubated at 37℃. Microbial and chemical changes were assessed during papaya fermentation and after four weeks of cold storage. Two selected isolates (Pw1 and Cb4) had in vitro cholesterol reduction of > 80%. These two isolates lowered lipid profile (triglyceride, total cholesterol, LDL-c) significantly, and increased HDL-c levels (p < 0.5) in the rat sera. Phylogenetic analysis showed that Pw1 was 98.86% similar to Limosilactobacillus fermentum, while Cb4 was 99.54% similar to Enteroccocus faecium. Both strains fermented papaya juice with cell viability reaching 8.92 × 108 cfu/ml and 25.3 × 108 cfu/ml respectively, and were still viable after 4 weeks of cold storage.

Exopolysaccharide (EPS) Production by Lactobacillus paracasei KLB58 in Modified Medium under Different Growth Conditions (다양한 배지 환경이 Lactobacillus paracasei KLB 58의 Exopolysaccharide (EPS) 생산량에 미치는 영향)

  • Lee, Choong-Young;Jeon, Jeong-Min;Lee, Hae-In;Kim, Min-Hee;Jung, Mi-Kyoung;So, Jae-Seong
    • KSBB Journal
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    • v.23 no.1
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    • pp.18-22
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    • 2008
  • Various probiotic Lactobacillus spp. are known to produce exopolysaccharide (EPS) which has potential health promoting functionality. A Lactobacillus paracasei strain producing EPS was isolated from healthy human. This strain, named L. paracasei KLB58, was grown on modified MRS medium. Experiments were conducted under various growth conditions to optimize the EPS production. Our study showed that incubation temperature played an important role in EPS production. When incubation temperature was changed from $37^{\circ}C$ to $25^{\circ}C$, the increase of EPS production (28.1 mg/ml) was the highest in our experiment. The type of carbon source in the medium also affected EPS production. Galactose was the most effective for EPS production among the carbon sources examined. Using galactose, glucose, lactose and sucrose, the amount of released EPS was 38.9 mg/ml, 35.6 mg/ml, 21.76 mg/ml and 16.9 mg/ml, respectively. However, acidity in growth medium inhibited EPS productivity due to the low growth yield. When grown at pH 4, L. paracasei KLB58 could only produce EPS of 14.6 mg/ml. When the initial amounts of nitrogen and carbon sources were examined, EPS production was not significantly affected by nitrogen source while carbon source affected considerably.

Evaluation of Antidepressant Drug Effect in a Depressive Animal Model by Proton MR Spectroscopy (양성자 자기공명분광법을 이용한 우울증 동물모델에서의 항우울제 약물 효능 평가)

  • Kim, Sang-Young;Choi, Chi-Bong;Lee, Sung-Ho;Woo, Dong-Cheol;Yoon, Seong-Ik;Hong, Kwan-Soo;Lee, Hyun-Sung;Cheong, Chae-Joon;Jee, Bo-Keun;Hong, Sung-Tak;Kim, Hwi-Yool;Choe, Bo-Young
    • Progress in Medical Physics
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    • v.19 no.2
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    • pp.95-101
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    • 2008
  • In this study, we observed the alteration of choline signal intensity in hippocampus region of the depressive rat model induced by forced swimming test (FST). The purpose of this study was to evaluate the antidepressant efficacy in the depressive animal model using MR spectroscopy. Fourteen experimentally naive male Sprague-Dawley rats weighting $160{\sim}180\;g$ were used as subjects. Drug injection group was exposed to the FST except for control group. The drugs were administered subcutaneously (SC) in a volume equivalent to 2ml/kg. And three injections were administered 23, 5, and 1h before beginning the given test. 1H MR spectra were obtained with use of a point resolved spectroscopy (PRESS) localization sequence performed according to the following parameters: repetition time, 2500 ms; echo time, 144 ms; 512 average; 2048 complex data points; voxel dimensions, $1.5{\times}2.5{\times}2.5\;mm^3$ ; acquisition time, 25min. There were no differences in NAA/Cr and Cho/Cr ratio between the right and the left hippocampus both normal control rats and antidepressant-injected rats. Also, no differences were observed in NAA/Cr and Cho/Cr ratio between the normal control rats and the antidepressant-injected rats both the right and the left hippocampus. In this study, we found the recovery of choline signals in the depressive animal model similar to normal control groups as injecting desipramine-HCl which was antidepressant causing anti-immobility effects. Thus, we demonstrated that MR spectroscopy was able to aid in evaluating the antidepressant effect of desipramine-HCl.

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Effects of Molasses Supplementation on the Quality of Galgeuntang Meal Silage and the Palatability in Korean Native Goats (당밀의 첨가가 갈근탕박 사일리지의 품질과 산양의 기호성에 미치는 영향)

  • Kim, Sung-Bok;Moon, Gye-Bong;Lee, Bong-Duk;Oh, Hong-Rock;Bae, Hyung-Chul;Lee, Soo-Kee
    • Korean Journal of Agricultural Science
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    • v.33 no.1
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    • pp.25-34
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    • 2006
  • Two experiments were conducted to investigate the effects of molasses supplementation to silage materials in ensiling Galgeuntang (herbal medicine) meal on the quality of silage, and its palatability in Korean native goats. In experiment I, molasses was added at the levels of 0, 0.5, and 1.0% to Galgeuntang meal with three replicates per treatment. In experiment II, three 1.5-yr-old Korean native goats were used to measure palatability. In experiment I, lactic acid contents in molasses treatments were significantly(p<0.05) higher, and pH and butyric acid contents were lower than those of non-molasses treatment. In addition, molasses treatment increased total microbial cell counts in MRS medium for lactobacillus, but decreased total microbial cell counts in PDA medium for fungi. Molasses supplementation to silage materials increased in vitro dry matter disappearance. Molasses supplementation tended to increase silage intake in Korean native goat (experiment II), but the difference was not significant(p>0.05). It is concluded that molasses supplementation to silage materials in ensiling herbal medicine meal could improve its preservability and palatability, the higher the better.

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Functional Characteristics of Enterococcus faecium SA5 and Its Potential in Conversion of Ginsenoside Rb1 in Ginseng (Enterococcus faecium SA5의 기능적 특성과 인삼 ginsenoside Rb1의 전환)

  • Kim, Eun-Ah;Renchinkhand, Gereltuya;Urgamal, Magsal;Park, Young W.;Nam, Myoung Soo
    • Journal of Life Science
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    • v.27 no.2
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    • pp.172-179
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    • 2017
  • The fermentation of Panax ginseng can yield many compounds from ginsenosides that have a wide variety of biological functions. Lactic acid bacteria (LAB) strains are capable of converting ginsenosides. The purposes of this study were to: (1) characterize Enterococcus faecium SA5, an isolated LAB from Mongolian mare milk, (2) identify the existence of extracellular ${\beta}$-glucosidase activity in the milk, and (3) ascertain if the ${\beta}$-glucosidase has the capacity of converting ginsenoside in Korean ginseng. The results revealed that E. faecium SA5 was acid-resistant, bile salt-resistant, and has antibiotic activities against 4 pathogenic microorganisms (Salmonella typhimurium KCTC 3216, Listeria monocytogenes KCTC 3710, Bacillus cereus KCTC 1012, Staphylococcus aureus KCTC 1621). In addition, E. faecium SA5 had tolerance against some antibiotics such as colistin, gentamycin and neomycin. It was also found that E. faecium SA5 possessed bile salt hydrolase activity, which could lower blood cholesterol level. When incubated in 10% (w/v) skim milk as a yogurt starter, E. faecium SA5 caused to decrease pH of the medium as well as increase in viable cell counts. Using TLC and HPLC analysis on the samples incubated in MRS broth, our study confirmed that E. faecium SA5 can produce ${\beta}$-glucosidase, which was capable of converting ginsenoside $Rb_1$ into new ginsenosides $Rg_3-s$ and $Rg_3-r$. It was concluded that E. faecium SA5 possessed a potential of probiotic activity, which could be applied to yogurt manufacture as well as ginsenoside conversion in ginseng.

Enhanced Production, Purification, and Partial Characterization of Lacticin BH5, a Kimchi Bacteriocin Produced by Lactococcus lactis BH5

  • Paik, Hyun-Dong;Hyun, Hyung-Hwan;Pyun, Yu-Ryang;Ahn, Cheol;Hur, Ji-Woon;Kim, Tae-Seok;Yeo, Ick-Hyun
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.53-60
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    • 2000
  • Strain BH5 was isolated from naturally fermented Kimchi and identified as a bacteriocin producer, which has bactericidal activity against Micrococcus flavus ATCC 10240. Strain BH5 was identified tentatively as Lactococcus lactis by the API test and some characteristics. Lactococcus lactis BH5 showed a broad spectrum of activity against most of the non-pathogenic and pathogenic microorganisms tested by the modified deferred method. The activity of lacticin BH5, named tentatively as the bacteriocin produced by Lactococcus lactis BH5, was detected at the mid-log growth phase, reached its maximum during the early stationary phase, and decreased after the late stationary phase. Lacticin BH5 also showed a relatively broad spectrum of activity against non-pathogenic and pathogenic microorganisms as tested by the spot-on-lawn method. Its antimicrobial activity on sensitive indicator cells was completely disappeared by protease XIV or ${\alpha}$-chymotrypsin. The inhibitory activities of lacticin BH5 were detected during treatments up to 100$^{\circ}C$ for 30 min. Lacticin BH5 was very stable over a pH range of 2.0 to 9.0 and was stable with all the organic solvents examined. The cell concentration and bacteriocin production in strain BH5 were maximum when grown at 30$^{\circ}C$ in a modified MRS medium supplemented with 0.5% tryptone, 1.0% yeast extract, and 0.5% beef extract as nitrogen sources. It demonstrated a typical bactericidal mode of inhibition against Micrococcus flavus ATCC 10240. Lacticin BH5 was purified through ammonium sulfate precipitation, ethanol precipitation, and CM-Sepharose column chromatography. The apparent molecular mass of lacticin BH5 was estimated to be in the region of 3.7 kDa, by the direct detection of bactericidal activity after SDS-PAGE. Mutant strain NO141 which was isolated by nitrosoguanidine mutagenesis produced about 4 fold more bacteriocin than the wild type.

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The Effect of Gamma Irradiation on the Acid Tolerance and Bile Tolerance of Lactic Acid Bacteria (감마선 조사가 젖산균의 내산성 및 내담즙성에 미치는 영향)

  • Kim, Jae-Kyung;Lee, Ji-hye;Park, Jong-Heum;Song, Beom-Seok;Lee, Ju-Woon;Choi, Jong-Il;Hwang, E-Nam;Kang, Sangmo;Park, Sang-Hyun;Kim, Jae-Hun
    • Journal of Radiation Industry
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    • v.6 no.2
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    • pp.177-180
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    • 2012
  • The acid and bile tolerance changes of 5 different lactic acid bacteria (LAB; Lactobacillus paracasei, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus debruekii) with gamma irradiation were evaluated. The abilities of LAB to survive in the acidic conditions at the stomach and the bile acidic conditions at the beginning of the small intestine are the key functions for applying LAB to probiotics. In the results, all of LAB survived more than 50% after incubation in PBS (pH 2.5) for 2 hr, which indicated more than half of LAB are possible to pass through the stomach. However, gamma irradiation decreased the acid tolerances of LAB. The bile tolerances of all bacteria except Lactobacillus acidophilus were observed to survive at a 3% oxgall concentration in MRS, and 1 kGy of gamma irradiation to LAB did not affect any bile tolerances changes. But gamma irradiated Lactobacillus casei and Lactobacillus casei (3 kGy) showed decreasing survival rate with oxgall added MAS agar. In conclusion, gamma irradiation should be applied to yogurt or fermented foods with care because LAB could be changes their properties on acid and bile tolerances.

Effects of Molasses Supplementation Levels to Daesihotang (Herbal Medicine) Meal on Quality of Silage and the Palatability in Goats (당밀의 첨가가 대시호탕(한약제)박 사일리지의 품질과 산양의 기호성에 미치는 영향)

  • 김성복;문계봉;이봉덕;배형철;이수기
    • Journal of Animal Science and Technology
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    • v.48 no.5
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    • pp.683-690
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    • 2006
  • Two experiments were conducted to investigate the effects of molasses supplementation levels to silage materials in ensiling herbal medicine meal on the quality of silage, and its palatability in Korean native goats. In experimentⅠ, molasses were added at the levels of 0, 0.5, and 1.0% to herbal medicine (Deasihotang) meal, with three replicates per each level. They were kept at room temperature for 40 d in glass bottles in order to get silage samples. In experiment Ⅱ, nine 1.5-yr-old Korean native female goats were employed to measure their palatability. In experimentⅠ, lactic acid contents in molasses treatments were significantly (p<0.05) higher, and pH and butyric acid contents was lower than those of non-molasses control treatment. In addition, molasses treatments increased total microbial cell counts in MRS medium for lactobacillus, but decreased total microbial cell counts in PDA medium for fungi. Molasses supplementation to silage materials increased in vitro dry matter disappearance rate. Molasses supplementation tended to increase silage intake in Korean native goat (experiment II), however, the difference was not significant. It is concluded that molasses supplementation to silage materials in ensiling Daesihotang meal could improve its preservability and palatability, the larger the amount the better the quality.

Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.519-526
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    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

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Improvement of Kimchi Fermentation by Using Acid-Tolerant Mutant of Leuconostoc mesenteroides and Aromatic Yeast Saccharomyces fermentati as Starters

  • Kim, Young-Chan;Jung, Eun-Youg;Kim, Hyung-Joo;Jung, Dai-Hyun;Hong, Seong-Gil;Kwon, Tae-Jong;Kang, Sang-Mo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.22-31
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    • 1999
  • Saccharomyces fermentati and Leuconostoc mesenteroides were isolated from a traditional kimchi, and then the Leu. mesenteroides was mutated to the acid-tolerant mutant Leu. mesenteroides M-l00. In the result of growth properties in MRS broth with various pHs adjusted with HCl and acid solution (latic acid:acetic acid=1:2), an acid-tolerant mutant Leu. mesenteroides M-100 showed more increased ability for growth than its wild strain at various temperatures. The strains were used as starters for the fermentation of kimchi. The experiments were performed with classified experimental groups (Group I, control kimchi; Group II, addition of YK-19 only; Group III, addition of M-100 only; Group IV, addition of mixture of M-100 and YK-19), and their pH, total acidity, reducing sugars content, organic acid productivity, organoleptic tests, and microfloral changes were compared. As a result, in pH and acidity, the optimal ripening period of Group IV was about 13.5 days (i.e. from the 8.5 to 22 days of fermentation). This result indicates that the optimal ripening period of Group IV was about 1.5 times longer than that of Group I. Furthermore, Group IV was edible to 28 days of fermentation. In addition, high contents of succinc acid was observed in Group IV. Group IV was also highly ranked on the organoleptic test. During the fermentation of kimchi, the number of Leuconostoc sp. in group I reduced after 7 days; however, the number of Leuconostoc sp. in Group II, III, and IV decresed after 14 days of fermentation. An especially high number of Leu. sp. was observed in Group IV, and this gave better flavor of kimchi than any other during the whole fermentation period. Citric acid, tartaric acid, succinic acid, fumaric acid, and lactic acid were detected in the kimchi, and a significant increase in the concentration of lactic acid during fermentation was observed in the all experimental groups.

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