• Archives of Pharmacal Research
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• v.11 no.3
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• pp.218-224
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• 1988

The preparation of Clostridium butyricum is used as a normalizing agent for human intestinal flora. When the microbe is simultaneously used with rifampicin, it is inactivated by the antibiotic. To develop rifampicin-resistant mutants, rifampicin-sensitive strain Miyairi II 588 of C. butyricum was treated with nitrosoguanidine (NTG). To ensure stable resistance to rifampicin, we examined whether the resistance was plasmid-mediated or chromosome-mediated. It was found that the resistance of four mutant strains was not mediated by its inherent plasmid, but by the chromosomal mutation. These strains were examined for the susceptibility and resistance to other antituberculosis agents and antibiotics. The results showed that these mutants were resistant to the high concentration of the antituberculosis agents.

• Journal of Animal Science and Technology
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• v.53 no.4
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• pp.341-348
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• 2011

The antagonistic activities against animal entero-pathogenic bacteria were investigated with 444 natural substances fermented by various probiotics. A white rice product fermented (FWR) by Clostridium butyricum IDCC 9207 with a high growth inhibition of Salmonella typhimurium KCTC 2054 and Escherichia coli O157:H7 was selected. Also, a FWR was shown to suppress 8 among 21 pathogenic bacteria. In a mouse model with salmonella (${\times}10^9$ CFU/mouse) infection, 5 samples (200 ${\mu}{\ell}$/mouse/day) were fed to mice (n = 25) for 18 days. A fermented white rice containing C. butyricum IDCC 9207 (FWRCb9207) among 5 samples significantly inhibited the growth of salmonella, while in the control group (PBS, tetracycline) the number of salmonella increased. And the treatment with FWRCb9207 was found to increase the secretory immunoglobulin A (sIgA) level in the feces of salmonella-infected mice. The results obtained in this study suggest that a FWRCb9207 might be utilized as a feed additive in pigs and poultry diets.

• Transactions of the Korean hydrogen and new energy society
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• v.15 no.4
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• pp.309-316
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• 2004

Chlamydomonas reinhardtii UTEX 90 was cultivated with continuous supply of 2% $CO_2$ using TAP media at $25^\circ{C}$ and produced biomass 1.18 g of dry cell weight/L for 4 days. C. reinhardtii algal biomass(CAB) was concentrated to 20 times by volume and converted into hydrogen and organic acids by anaerobic fermentation using Clostridium butyricum. Organic acids in the fermentate of CAB were consecutively used to produce hydrogen by Rhodobacter sphaeroides KD 131 under the light condition. Approximately 52% of starch in the concentrated CAB which had 4-5.8, 24-26 and 6-7 g/L of starch, protein and fat, respectively was degraded by Cl. butyricum at $37^\circ{C}$. During this process, hydrogen and some organic acids, such as formate, acetate, propionate, and butyrate, respectively were produced. Further conversion of the organic acids in anaerobic fermentate of CAB by Rb. sphaeroides KD131 produced hydrogen from the anaerobic fermentate under the illumination of 8 klux using halogen lamp at $30^\circ{C}$. The result showed that hydrogen was evolved by the anaerobic conversion using Cl. butyricum and then by the photosynthetic fermentation using Rb. sphaeroides KD131. It indicated that the two-step conversion process produced the maximum amount of hydrogen from algal biomass which contained carbohydrate, protein, and fat via organic acids.

• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.56-62
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• 1989

To find bacteria which can inhibit growth of enteropathogenic Clostridium perfringens ATCC 13124, spore forming butyric acid bacteria were isolated from 26 fecal samples of infants. Fourteen strains were found to be antagonistic to the enteropathogen and five of them produced butyric acid. A strain which produced the highest butyric acid was selected and identified as Clostridium butyricum. This organism sporulated in SM medium in 36 hours with optimum rates at 37$^{\circ}C$ and at pH 5.5. The spores tolerated well at high heat and acidity, and possible application of Clostridium butyricum as intestinal controller was discussed.

• Journal of Life Science
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• v.12 no.5
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• pp.595-604
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• 2002

The growth promoting effect of Clostridium hutyricum KCTC 1785 was investigated with natural products bearing antioxidative capacity, and combined two, three and four kinds of them. C. butyricum was showed a good growth by Lycii fructus, Sophorae flos, Chelidonium majus L., Atractylodis rhizoma alba, Paeonia japonica, alone, and two mixed com-binations were composed of Paeonia japonica and Epimedii herba, Paeonia japonica and Aurantii nobilis pericarpium, Paeonia japonica and Puerariae radix, Pneonia japonica and Angelicae gigantis radix, and three mixed combinations were organized with Epimedii herba, Sophorae flos and Nnelumbo nuclfera gaertner, and Epimedii herba, Sophorae flos and Scutellaria haicalensis george, and Epimedii herba, Sophorae flos and theae folium, and Epimedii herba, Paeonia japonica and Angelicae gigantis radix, and four mixed combinations were formed with Epimedii herba, Puerariae radix, Nelumbo nuclfera gaertner and Paeonia japonica, and Epimedii herba, Puerariae radix, Nelumbo nuclfera gaertner and Theae folium, and Epimedii berba, puerariae radix, Nelumbo nuclfera gaertner and Angelicae gigantis radix, and puerariae radix, Nelumbo nuclfera gaertner, paeonia japonica and Theae folium. As these combinations of natural products could activate some parts fo body, they might be applied to pharmaceuitcal applications, functional foods, antiaging tea, also expected to promote useful enterobacterial growth for multifunctional fermentative beverage.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.9
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• pp.1016-1021
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• 2005

We studied on screening and isolation of dominant bacteria in the food waste fermentation process and on effective production of VFAs by isolated bacteria. In the result of study, bacteria of twelve species were isolated by anaerobic medium. Among the 12 isolated species including Escherichia coli, Clostridium formicoaceticum, C. butyricum, C. acetobutyricum. E. coli and Clostridium spp. were occurred dominantly in the fermentation process and regarded as best VFAs producing bacteria. Acetic acid are produced 287 mg/gTS(8,176 mg/L) by E. coli in concentration of $6{\times}10^8\;cells/gTS$, 551 mg/gTS(15,715 mg/L) by Clostridium formicoaceticum in concentration of $5{\times}10^4\;cells/gTS$. Three times as much acetic acid were produced as blank. Butyric acid are produced 214 mg/gTS(6,106 mg/L) by C. butyricum in concentration of $2.5{\times}10^5\;cells/gTS$ and produced 254 mg/gTS(7,261 mg/L) by C. acetobutyricum of concentration of $1.5{\times}10^5\;cells/gTS$. Two times as much butyric acid were produced as blank.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.841-844
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• 2001

Free radicals cause many diseases and aging. In our body coexist useful and harmful bacteria. Some useful intestinal bacteria such as L. plantarum, B. adolescentis, B .bifidum, B. infantis, C. butyricum were presented positive growth in culture medium bearing natural products. L. plantarum was more effectively grown by mixed natural products of CORNI FRUCTUS+THEAE FOLIUM+COPTIDIS RHIZOMA+PAEONIA JAPONICA, B. adolescentis was BEIAMCANDA CHINENSIS+ANGELICAE GIGANTIS RADIX+EPIMEDII HERBA+PAEONIA JAPONICA, B. bifidum was NELUMBO NUCLFERA GAERTNER+THEAE FOLIUM+BEIAMCANDA CHINENSIS+CORNI FRUCTUS, B. infantis was THEAE FOLIUM+CHELIDONIUM MAJUS L.+ASTRAGALI RADIX+EPIMEDII HERBA, C. butyricum was EPIMEDII HERBA+PUERARIAE RADIX+NELUMBO NUCLFERA GAERTNER+PAEONIA JAPONICA,etc. These natural products will support an important role in bio-industry.

• Journal of Life Science
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• v.26 no.3
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• pp.353-358
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• 2016

Thiolase is an enzyme that catalyzes condensation reactions between two acetyl-CoA molecules to produce acetoacetyl-CoA. As thiolase catalyzes is the first reaction in the production of n-butanol, knowledge of the molecular and regulatory mechanism of the enzyme is crucial for synthesizing high-value biofuel. Thiolase from Clostridium butyricum (CbTHL) was expressed, purified, and crystallized. X-ray diffraction data were collected from the crystals, and the 3-dimentional structure of the enzyme was determined at 2.0 Å. The overall structure of thiolase was similar to that of type II biosynthetic thiolases, such as thiolase from C. acetobutylicum (CaTHL). The superposition of this structure with that of CaTHL complexed with CoA revealed the residues that comprise the catalytic and substrate binding sites of CbTHL. The catalytic site of CbTHL contains three conserved residues, Cys88, His349, and Cys379, which may function as a covalent nucleophile, general base, and second nucleophile, respectively. For substrate binding, the way in which CbTHL stabilized the ADP moiety of CoA was unlike that of other thiolases, whereas the stabilization of β-mercaptoethyamine and pantothenic acid moieties of CoA was quite similar to that of other enzymes. The most interesting observation in the CbTHL structure was that the enzyme was regulated through redox-switch modulation, using a reversible disulfide bond.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1636-1643
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• 2014

3-Hydroxybutyryl-CoA dehydrogenase is an enzyme that catalyzes the second step in the biosynthesis of n-butanol from acetyl-CoA, in which acetoacetyl-CoA is reduced to 3-hydroxybutyryl-CoA. To understand the molecular mechanisms of n-butanol biosynthesis, we determined the crystal structure of 3-hydroxybutyryl-CoA dehydrogenase from Clostridium butyricum (CbHBD). The monomer structure of CbHBD exhibits a two-domain topology, with N- and C-terminal domains, and the dimerization of the enzyme was mostly constituted at the C-terminal domain. The mode of cofactor binding to CbHBD was elucidated by determining the crystal structure of the enzyme in complex with $NAD^+$. We also determined the enzyme's structure in complex with its acetoacetyl-CoA substrate, revealing that the adenosine diphosphate moiety was not highly stabilized compared with the remainder of the acetoacetyl-CoA molecule. Using this structural information, we performed a series of site-directed mutagenesis experiments on the enzyme, such as changing residues located near the substrate-binding site, and finally developed a highly efficient CbHBD K50A/K54A/L232Y triple mutant enzyme that exhibited approximately 5-fold higher enzyme activity than did the wild type. The increased enzyme activity of the mutant was confirmed by enzyme kinetic measurements. The highly efficient mutant enzyme should be useful for increasing the production rate of n-butanol.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.111-118
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• 1988

The growth inhibition of enteropathogenic Escheriohia coli $A_2$and Escherichia coli G$_7$, causing the diarrhea in piglets, by the organic acid producing bacteria was studied in vitro. The metabolites of the organic acid bacteria, such as lactic acid, acetic acid inhibited the growth of E. coli $A_2$and E. coli G$_7$ in BL medium. The more the organic acid producing bacteria have ability to produce the organic acids, the higher these bacteria excelled the inhibitory efficacy against enteropathogenic E. coli. Among the strains examined, Lactobacillus casei Y and Streptococcus faecium C showed relatively strong growth inhibition against enteropathogenic E. coli.. When the organic acid producing bacteria and the enteropathogenic E. coli were incubated simultaneously in BL medium, bacteriostasis of E. coli was observed when the pH of BL culture was lowered to 5.0, and bacteriocidal effect was observed when the pH became Bess than 4.5, E. coli. $A_2$was more resistant to the organic acid bacteria than E. coli G$_7$.