• Microbiology and Biotechnology Letters
• /
• v.17 no.1
• /
• pp.63-68
• /
• 1989

The optimum conditions for cellular autolysis in Clostridium butyricum ID-113 have been investigated. Cellular autolysis was optimal at pH 1.0 in 0.05 M potassium phosphate buffer and at 37$^{\circ}C$. The rate of cellular autolysis depended on the age of culture. The most rapid cellular autolysis occurred in the cells of mid-exponentially growing cultures, but cellular autolysis decreased sharply when the cultures entered the stationary phase. A growing culture of Cl. butyricum ID-113 was induced to autolyze and lost its turbidity spontaneously in the hypertonic NaCl, sucrose, or glucose medium. The autolytic enzyme activity was found In the autolysate of cells and the supernatant of the culture.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.3
• /
• pp.368-377
• /
• 2020

Enterotoxigenic Bacteroides fragilis (ETBF) is the main pathogen causing severe inflammatory diseases and colorectal cancer. Its biofilm plays a key role in the development of colorectal cancer. The objective of this study was to determine the antagonistic effects of cell-free supernatants (CFS) derived from Clostridium butyricum against the growth and biofilm of ETBF. Our data showed that C. butyricum CFS inhibited the growth of B. fragilis in planktonic culture. In addition, C. butyricum CFS exhibited an antibiofilm effect by inhibiting biofilm development, disassembling preformed biofilms and reducing the metabolic activity of cells in biofilms. Using confocal laser scanning microscopy, we found that C. butyricum CFS significantly suppressed the proteins and extracellular nucleic acids among the basic biofilm components. Furthermore, C. butyricum CFS significantly downregulated the expression of virulence- and efflux pump-related genes including ompA and bmeB3 in B. fragilis. Our findings suggest that C. butyricum can be used as biotherapeutic agent by inhibiting the growth and biofilm of ETBF.

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

• Microbiology and Biotechnology Letters
• /
• v.17 no.1
• /
• pp.69-73
• /
• 1989

Cellular autolytic enzyme was isolated from the supernatant fluid of exponentially growing cuiture of Cl. butyricum ID-113. The autolysin was partially pruified by ammonium sulfate fractionation, chromatography on DEAE-Sephadex A-50 and gel filtration through Sephadex G-200. This autolytic enzyme lysed SDS-treated cell wall fractions of Cl. butyricum ID, but not whole cells at all. Its optimum pH and temperature were 5.0 and 37$^{\circ}C$, respectively. This enzyme was relatively stable at neutral pH, but sensitive to heat treatment. Enzyme activity was not influenced by the addition of various divalent cation, but inhibited by Cu$^{++}$.

• Korean Journal of Food Science and Technology
• /
• v.53 no.6
• /
• pp.775-784
• /
• 2021

After isolating the DIMO 52 strain with a large inhibition zone diameter for Clostridium perfringens and maximum butyric acid production from the fecal sample of a breastfeeding infant, it was identified as Clostidium butyricum. The maximum growth of the DIMO 52 strain was reached 24 h after inoculation, and the maximum butyric acid concentration was approximately 34.73±4.27 mM. The DIMO 52 strain survived approximately 67.5% of the initial inoculum at pH 2.0, and approximately 64.9% survived in RCM broth supplemented with 0.3% (w/v) oxgall. In addition, DIMO 52 showed antibacterial activity against Escherichia coli KCTC 2441 and Salmonella Typhimurium KCTC 1925. In LPS-stimulated RAW264.7 cells, 1×10³ CFU/mL viable cells of the DIMO 52 strain also exhibited significant NO (nitric oxide) production inhibitory activity (33%, p<0.01). This result suggests that C. butyricum DIMO 52 has anti-inflammatory activity related to NO radical-scavenging activity. In conclusion, C. butyricum DIMO 52 isolated in this study has the potential to be used as a probiotic.

• Archives of Pharmacal Research
• /
• v.11 no.3
• /
• pp.218-224
• /
• 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 Korean Society of Environmental Engineers
• /
• v.27 no.9
• /
• pp.1016-1021
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.8
• /
• pp.1210-1215
• /
• 2006

The properties related to the temperature and oxygen stability of the cytoplasmic hydrogenases from the fermentative strict anaerobic bacterium, Clostridium butyricum NCIB 9576 (Cl. butyricum), and purple sulfur phototrophic bacterium, Thiocapsa roseopersicina NCIB 8347 (T. roseopersicina), were compared. The optimum temperatures for the growth of Cl. butyricum and T. roseopersicina were 37$^{\circ}C$ and 25$^{\circ}C$, respectively, whereas those for the H$_2$ evolution of the cytoplasmic hydrogenases prepared from Cl. butyricum (C-H$_2$ase) and T. roseopersicina (T-H$_2$ase) were 45$^{\circ}C$ and 65$^{\circ}C$, respectively. The T-H$_2$ase was more thermostable than the C-H$_2$ase and retained its full activity for 5 h at 50$^{\circ}C$ under anaerobic conditions and 90% of its activity at 60$^{\circ}C$, whereas the C-H$_2$ase lost its activity drastically at 50$^{\circ}C$. The optimum pHs for H$_2$ oxidation of the C-H$_2$ase and T-H$_2$ase were 9.0 and 7.5, respectively. Both enzymes showed a maximum H$_2$ evolution activity at pH 7.0. Under aerobic conditions, 80% of the T-H$_2$ase activity was retained for 10 h at 30$^{\circ}C$, and 50% of the activity remained after 6 days under the same experimental conditions. However, the C-H$_2$ase was labile to oxygen and lost its activity immediately on exposure to air. Therefore, these properties of the T-H$_2$ase are expected to be advantageous for application in in vitro biological H$_2$ production systems.

• Microbiology and Biotechnology Letters
• /
• v.27 no.1
• /
• pp.62-69
• /
• 1999

Clostridium butyricum NCIB 9576 evolved hydrogen gas and produced various organic acids from glucose, lactose, starch, and glycerol. Total amount of hydrogen gas produced from 1 and 2% glucose were 630 and 950ml $H_2$/l-broth, respectively, for the first 24 hrs of incubation and the maximum hydrogen production rates were 42 and 94ml $H_2$/hr/1-broth, respectively. Teh initial pH 6.8 decreased to 4.2~4.5 during the first 12~16 hrs of fermentation when the pH was not controlled, resulting in ceasing the cell growth and hydrogen evolution and in degradation of 82 and 40% glucose after 24hrs of incubation from 1 and 2% glucose, respectively. When pH was controlled to 5.5, glucose was consumed completely and resulted in increasing hydrogen production approximately 38~50% compared to the experiments without the pH control. C. butyricum NCIB 9576 produced hydrogen gas approximately 644, 1,700 and 3,080 ml $H_2$/l-broth with 0.5, 1 and 2% lactose, respectively and the maximum hydrogen production rates were 41, 141 and 179ml $H_2$/hr/l-broth, respectively. All of the lactose added was degraded completely during fermentation even though pH was not controlled. C. butyricum NCIB 9576 produced 183 and 709ml $H_2$/l-broth with 0.1 and 0.5% starch for 48 hrs, respectively, when pH was not controlled. The maximum rates of hydrogen gas production were 43 and 186ml $H_2$/l-broth, respectively and 80~100% of starch added was fermented. Approximately 107ml $H_2$/l-broth was produced using 1% glycerol by C. butyricum NCIB 9576 and the pH was maintained higher than 6.1 during fermentation without pH control. The degradation of glucose, lactose, starch and glycerol by C. butyricum NCIB 9576 were affected by the pH of fermentation broth and the organic acids released during fermentation. The pH of feremtntation broth dropped to 4.2~4.6 after 12~14 hrs incubation when glucose was used as a substrate while pHs were maintained above pH 5 under the same experimental conditions when lactose, starch and glycerol were used. The organic solvents and acids produced during glucose fermentation were mainly ethanol, butyrate, acetate and a little of propionate, while butyrate was the main organic acids during the lactose, starch, and glycerol fermentation by C. butyricum NCIB 9576.

• Journal of Microbiology and Biotechnology
• /
• v.33 no.10
• /
• pp.1337-1350
• /
• 2023

Caproic acid is a precursor substance for the synthesis of ethyl caproate, the main flavor substance of nongxiangxing baijiu liquor. In this study, Clostridium butyricum GD1-1, a strain with high caproic acid concentration (3.86 g/l), was isolated from the storage pit mud of nongxiangxing baijiu for sequencing and analysis. The strain's genome was 3,840,048 bp in length with 4,050 open reading frames. In addition, virulence factor annotation analysis showed C. butyricum GD1-1 to be safe at the genetic level. However, the annotation results using the Kyoto Encyclopedia of Genes and Genomes Automatic Annotation Server predicted a deficiency in the strain's synthesis of alanine, methionine, and biotin. These results were confirmed by essential nutrient factor validation experiments. Furthermore, the optimized medium conditions for caproic acid concentration by strain GD1-1 were (g/l): glucose 30, NaCl 5, yeast extract 10, peptone 10, beef paste 10, sodium acetate 11, L-cysteine 0.6, biotin 0.004, starch 2, and 2.0% ethanol. The optimized fermentation conditions for caproic acid production by C. butyricum GD1-1 on a single-factor basis were: 5% inoculum volume, 35℃, pH 7, and 90% loading volume. Under optimal conditions, the caproic acid concentration of strain GD1-1 reached 5.42 g/l, which was 1.40 times higher than the initial concentration. C. butyricum GD1-1 could be further used in caproic acid production, NXXB pit mud strengthening and maintenance, and artificial pit mud preparation.