• Journal of Microbiology and Biotechnology
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• 제2권4호
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• pp.293-296
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• 1992

Increased concentrations of yeast extract led to increased growth yields and faster growth rates of the newly isolated Clostridium thermobutyricum. This species produced butyrate as its main fermentation product from glucose as well as from yeast extract. In the presence of peptone or tyrptone and during growth on agar, up to 70% of the cells sporulated. Growth yields were 30 and 55 g per mole glucose in the presence of 0.05 and 2.0% yeast extract, respectively. The Arrhenius graph was biphasic, exhibiting an intermediary plateau around $38^{\circ}C$ with a concomitant change in the Arrhenius energy. The optimum temperature was $55^{\circ}C$. An unusually sharp decline in the growth rate occurred above $59^{\circ}C$ .

• 대한기계학회:학술대회논문집
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• 대한기계학회 2008년도 추계학술대회A
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• pp.1643-1646
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• 2008

In this study, an elastic network model is established in order to find dominant factors which reflect thermostability of protein structures. The connections in the elastic network model are selected with respect to the free energy between alpha-carbons, which is representatives of residues in the elastic network model. We carried out normal mode analysis and compared eigenvalues of the stiffness matrix from the elastic network model. In most cases, thermophilic proteins are observed to have higher values of lowest natural frequency than mesophiles and psychrophiles have. As a result, the thermophiles are calculated to be stiffer than other proteins in view of dynamic vibration.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.165-168
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• 2001

Regulation of pyrimidine nucleotide synthesis has been studied extensively in enteric bacteria and Bacillus species. Varieties of control modes have been proposed for regulation of pyrimidine nucleotide biosynthetic (pyr) genes. In Bacillus caldolyticus and B. subtilis, it has been proved that pyrimidine de novo biosynthetic operon is controlled by a regulatory protein PyrR-mediated attenuation. Another Gram-positive bacteria including Enterococcus faecalis, Lactobacillus plantarum, and wctococcus lactis have been found to constitute a pyr gene cluster containing the pyrR gene. In addition, it has been proposed that the structure of the 5' leader region of the Gram-negative extreme thermophile Thermus strain Z05 pyr operon provides a novel mechanism of PyrR-dependent coupled transcription-translation attenuation. Bacterial genome sequencing projects have identified the PyrR homologues in Haemophilus influenzae, Synechocystis sp., Mycobacterium tuberculosis, Streptococcus pneumoniae, S. pyogenes, and Clostridium acetobutylicum, which are currently investigating for their physiological functions.

• Journal of Life Science
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• 제11권2호
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• pp.120-125
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• 2001

The pyrR gene of the pyrimidine biosynthesis (pyr) operon of the thermophile Bacillus caldolyticus, encoding a uracil phosphoribosyltransferase (UPRTase), turned to rely as a pyr operon regulator. It has been proposed that PyrR mediates transcriptional termination-antitermination at three intercistronic regions of the par operon (S.-Y Ghim and J. Neuhard, J. Bacteriol.,176, 3698-3707, 1994). In this research, a plasmid carrying the pyrR region of B. caldolyticus could restore a pyrimidine regulation in a pyrR mutant of B. subtilis. Expression of pyrR was found to increase 6-7 fold during pyrimidine starvation. Additionally, a highly conserved nucleotide sequence which may constitute the binding site for a PyrR protein (PyrR-binding loop) in transcript was staggested. Alternative antiterminator and terminator structures involving three conserved motifs in front of the pyrR, pyrP and pyrB genes, respectively, are proposed to account for the observed regulation pattern.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.695-703
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• 2008

A locally isolated thermophile, Geobacillus sp. SAB-40, producing thermostable extracellular amylase constitutively and an induced intracellular ${\beta}$-galactosidase was characterized and identified based on 16S rRNA sequencing. A phylogenetic analysis then revealed its closeness to Geobacillus stearothermophilus. To evaluate the effect of the culture conditions on the coproduction of both enzymes by G stearothermophilus SAB-40, a Plackett-Burman fractional factorial design was applied to determine the impact of twenty variables. Among the tested variables, $CaCI_2$, the incubation time, $MgSO_4{\cdot}7H_2O$, and tryptone were found to be the most significant for encouraging amylase production. Lactose was found to promote ${\beta}$-galactosidase production, whereas starch had a significantly negative effect on lactase production. Based on a statistical analysis, a preoptimized medium attained the maximum production of amylase and ${\beta}$-galactosidase at 23.29 U/ml/ min and 12,958 U/mg biomass, respectively, which was 3-and 2-fold higher than the yield of amylase and lactase obtained with the basal medium, respectively.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.611-615
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• 2008

Novel groups of uncultivable anaerobic thermophiles were isolated from compost by enrichment cultivation in medium with a cell-free extract of Geobacillus toebii. The cell-free extract of G. toebii provided the medium with growth-supporting factors (GSF) needed to cultivate the previously uncultured microorganisms. Twenty-nine GSF-requiring candidates were successfully cultivated, and 16 isolated novel bacterial strains were classified into three different groups of uncultivable bacteria. The similarity among these 16 isolates and a phylogenetic analysis using 16S rRNA gene sequences revealed that these GSF-requiring strains represented novel groups within the family Clostridiaceae.

• Applied Microscopy
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• 제30권4호
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• pp.413-421
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• 2000

초고온 archaeon인 Thermococcus profundus에서 실린드형태의 단백질 복합체를 분리, 생화학적 특성과 구조를 규명하였다. 전자현미경과 영상처리 (image processing)를 이용한 구조 분석으로 8개의 subunit으로 이루어진 링 (ring) 형태가 두 개 겹쳐져 가운데 통로를 가지는 실린드형태로 이루어진 복합체임을 알 수 있었다. 또한 60kDa단백질(P60 complex)로 구성된 homomultimer임을 보여 주고 있다. 이 복합체는 $80^{\circ}C$ 이상에서 도 강한 ATPase의 활성을 보여주는 강한 내열성 단백질임을 알 수 있다. P60 complex는 초고온 archaeon이 높은 온도에서 적응하며 생존하는데 매우 중요한 역할을 할 것으로 추정할 수 있다.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.469-474
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• 1999

Thermoactinomyces sp. E79 produced two types of thermostable alkaline proteases extracellularly. A minor protease was separated from a major protease by using DEAE-column chromatography. This enzyme was purified to homogeneity by ammonium sulfate and DEAE-Sepharose ion-exchange chromatography. The purified minor protease showed different biochemical properties compared to the major protease. The molecular mass of the purified enzyme was estimated by SDS-PAGE to be 36 kDa. Its optimum temperature and pH for proteolytic activity against Hammarsten casein were $70^{\circ}C$ and 9.0, respectively. The enzyme was stable up to$75^{\circ}C$ and in an alkaline pH range of 9.0-11.0. The enzyme was inhibited by phenylmethylsulfonyl fluoride (PMSF) and $Hg^{2+}, indicating that the enzyme may be a cysteine-dependent serine protease. In addition, the enzyme cleaved the endoproteinase substrate, succinyl-Ala-Ala-Pro-Phe-p- nitroanilide, and the $K_m$ value for the substrate was 1.2 mM.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.1008-1012
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• 2003

Symbiobacterium toebii has been reported as a thermophile exhibiting a commensal interaction with Geobacillus toebii. The distribution of the commensal thermophiles in various soils was investigated using a denaturing gradient gel electrophoresis (DGGE). Based on the DGGE analysis, the enrichment condition for the growth of Symbiobacterium sp. was found to also enrich populations of several other microbial spp. as well as Symbiobacterium sp. In the enrichment experiment, several different 16S rDNA sequences of commensal thermophiles were detected in all of the soil samples tested, indicating that commensal thermophiles are widely distributed in various soils.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1986년도 추계학술대회
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• pp.515.1-515
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• 1986

Thermostable tryptophanase was extracted from a thermophilie bacterium, strain T which was absolutely symbiotic with strain 5. The enzyme was purified 14.7 fold with 5.8% yield by chromatographies using ion exchange, gel filtration, and hydrophobic interaction columns, followed by high performance liquid chromatography on hydroxyapatite column. The purified enzyme has a molecular weight of approximately 210,000 estimated by gel filtration column chromatography, and the molecular weight of subunit was determined by SDS polyacrylamide gel electrophoresis to be 46,000, which indicates that the native enzyme is made of four homologous subunits. The tryptophanase was stable at 65o0 and the optimum temperature for the enzyme activity for 20 min reaction was 70$^{\circ}C$. The purified enzyme activity for 20 min ieaction was 70$^{\circ}C$. The purified enzyme catalyzed the degradation of L-tryptophan into indole, pyruvate and ammonia in the presence of pyridoxal phosphate. 5-Hydroxy-Ltryptophan, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-L-cysteine, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-Lcysteine, and L-serine were also used as substrates to form pyruvate. The amino acid composition of the tryptophanase was determined, and found to contain a high percentage of hydrophobic amino acids, especially in the proline content, which was much higher than that of Escherichia coli tryptophanase. In addition, the 35N-terminal amino acid sequence of the tryptophanase was completely different from that of E. coli tryptophanase.