• KSBB Journal
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• v.5 no.1
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• pp.9-17
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• 1990

The growth and fermentation profiles of Clostridium acetobutylicum KCTC 1037 were examined in batch and continuous modes with pH variation and phosphate limitation. Clostridium acetobutylicum KCTC 10 37 grew better at pH 4.5 than at pH 5.5 or 6.5. Acetate and butyrate were produced at pH 5.5, whereas culture at pH 4.5 produced acetone and butanol. Solvent production was increased by the phosphate limitation in a batch culture, but in a phosphate-limited continuous culture for 400 hours steady-state solventogenesis was not observed. The induction and maintenance of solventogenesis presumably require not only acidic condition or phosphate limitation but also favourable bioenergetic condition.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.5
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• pp.432-443
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• 2005

Microarray technology has contributed Significantly to the understanding of bacterial genetics and transcriptional regulation. One neglected aspect of this technology has been optimization of microarray-generated signals and quality of generated information. Full genome microarrays were developed for Clostridium acetobutylicum through spotting of PCR products that were designed with minimal homology with all other genes within the genome. Using statistical analyses it is demonstrated that Signal quality is significantly improved by increasing the hybridization volume. possibly increasing the effective number of transcripts available to bind to a given spot, while changes in labeled probe amounts were found to be less sensitive to improving signal quality. In addition to Q-RT-PCR, array validation was tested by examining the transcriptional program of a mutant (M5) strain lacking the pSOL1 178-gene megaplasmid relative to the wildtype (WT) strain. Under optimal conditions, it is demonstrated that the fraction of false positive genes is 1% when considering differentially expressed genes and 7% when considering all genes with signal above background. To enhance genomic-scale understanding of organismal physiology, using data from these microarrays we estimated that $40{\sim}55%$ of the C. acetobutylicum genome is expressed at any time during batch culture, similar to estimates made for Bacillus subtilis.

• Korean Chemical Engineering Research
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• v.49 no.1
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• pp.105-108
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• 2011

The increased concern for the security of the oil supply and the negative impact of fossil fuels on the environment, particularly greenhouse gas emissions, has put pressure on society to find renewable fuel alternatives. Compared to the traditional biofuel, ethanol, higher alcohols offer advantage as gasoline substitutes because of their higher energy density and lower hygroscopicity. For this reason, microbial fermentation is known as potential producers for sustainable energy carriers. In this study, bacterial responses including cellular and molecular toxicity were studied in three different microorganisms, such as Escherichia coli, Clostridium acetobutylicum, and Saccharomyces cerevisiae. In this study, it was analyzed specific stress responses caused by ethanol and buthanol using four different stress responsive genes, i.e. fabA, grpE, katG and recA. The expression levels of these genes were quantified by semi-quantitative reverse transcription-PCR. It was found that four genes have shown different responsive patterns when E. coli cultures were under stressful conditions caused by ethanol and buthanol, respectively. Therefore, in this study, the stress responsive effects caused by these alcohols and the extent of each stress response can be analyzed using the expression levels and patterns of different stress responsive genes.

• Applied Chemistry for Engineering
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• v.22 no.5
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• pp.447-452
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• 2011

Depletion of petroleum increased the need of alternative energy and chemical resources. Biomass, a renewable resource, can be transformed to bioenergy and biomaterials, and the materials from biomass will ultimately substitute petroleum based energy and chemical compounds. In this perspective, production of C4-C6 compounds for bioenergy and biomaterials are described for understating of current research progress. n-Butanol and n-butyric acid, the major C4 compounds, are produced by Clostridium tyrobutyricum, Clostridium beijerinckii, and Clostridium acetobutylicum. n-Hexanoic acid, a typical C6 compound, is produced by Clostridium kluyveri and Megasphaera elsdenii. Reported maximum amount of n-butanol, n-butyric acid and n-hexanoic acid was 21, 55, and 19 g/L, respectively, and extraction of these C4-C6 compounds are induced increase production by those anaerobic bacteria. In addition, a new bacterium Clostridium sp. BS-1 produced 5 g/L of n-hexanoic acid using galactitol.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.85-91
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• 2001

To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

• Korean Journal of Veterinary Service
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• v.37 no.3
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• pp.157-164
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• 2014

Clostridium chauvoei is the etiologic agent of blackleg, a high mortality rated disease infection mainly cattle. In the present study, the partial sequences of 16S rRNA and flagellin gene of C. chauvoei isolated in Jeonbuk, Korea were determined and compared with those of reference strain. Oligonucleotide primers were designed to amplify a 811 bp fragment of 16S rRNA gene and 1229 bp fragment of flagellin gene. Sequencing analysis of 16S rRNA gene showed high homology to the reference strains ranging 82.3% to 100%, while flagellin gene were different from published foreign clostridia, showing 98.7% to 72.0% nucleotide sequence homology. Phylogenetic analysis based on 16S rRNA gene revealed the close phylogenetic relationship of C. chauvoei and C. septicum in cluster I, which includes C. carnis, C. tertium, C. quinii, C. celatum, C. perfringens, C. absonum, C. botulinum B. Phylogentic analysis also revealed that flagellin gene formed a single cluster with C. chauvoei, C. septicum, C. novyi A, C. novyi B, C. tyrobutylicum, C. acetobutylicum. The genetic informations obtained from this study could be useful for the molecular study of C. chauvoei.

• KSBB Journal
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• v.32 no.3
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• pp.169-173
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• 2017

Butyric acid (C4 carboxylic acid) is used as an important compound in food, pharmaceutical, and chemical industries. Currently, butyric acid is mainly produced at the industrial scale through the petrochemical processes. Bio-based butyric acid has also gained attention, because the consumer prefers the food and pharmaceutical ingredients that are produced through fermentation. Clostridia is one of the well-known butyric acid producers, and massively engineered for enhanced production of butyric acid. In this paper, we reviewed the metabolic pathway of clostridia, especially Clostridium acetobutylicum and Clostridium tyrobutyricum, and summarized the metabolic engineering strategies of the strains for enhanced production of butyric acid.

• 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.

• Korean Journal of Soil Science and Fertilizer
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• v.34 no.5
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• pp.333-341
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• 2001

The Compost-decomposing-bacteria was isolated from livestock compost containing sawdust. The isolated bacteria was identified as Bacillus subtilis LYH201 by the method of the composition of the fatty acid with MIDI system and Bergey's manual. Cloning of CMCase encoding gene was accompanied by shotgun method. The pLK100 have yellow activity ring on CMC medium, that was carried 2.2 kb insert DNA in pBluescript II $SK^+$ vector, named BglC gene. The BglC was very similar to Pectobacterium carotovorum Gun_CLOAB(P15704) with score of 57% identity and 71% homology over 508 aa. The BglC was measured molecular weight 56 kDa by CMC-SDS-PAGE. Optimum cellulase activity Bacillus subtilis LYH201 was temperature $50^{\circ}C$ and pH 7.