• Title/Summary/Keyword: thermophile

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Estimation of Distribution of a Commensal Thermophile in Soil by Competitive Quantitative PCR and Terminal Restriction Fragment Length Polymorphism Analysis

  • Rhee, Sung-Keun;Hong, Seung-Pyo;Bae, Jin-Woo;Jeon, Che-Ok;Lee, Seung-Goo;Song, Jae-Jun;Poo, Ha-Ryoung;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.940-945
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    • 2001
  • Symbiobacterium toebii has been previously reported as a novel commensal thermophile exhibiting a commensal interaction with thermophilic Geobacillus sp. SK-1. We investigated the distribution of this commensal thermophile in various soils using molecular methods, such as quantitative PCR and terminal restriction fragment polymorphism analysis. Based on a nested competitive quantitative PCR the 16S rDNA of the commensal thermophile was only detected in compost soils at about $1.0{\times}10^4$ cpoies per gram of soil, corresponding to $0.25{\times}10^4$ cells per gram of soil. However, in an enrichment experiment at $60^{\circ}C$, about $1.0{\times}10^8$ copies of 16S rDNA molecules were detected per ml of enriched culture broth for all the soils, and more than 0.1 mM indole accumulated as the product of commensal bacterial growth. When incubated at $30^{\circ}C$, neither the 16S rDNA of the commensal bacterium nor any indole accumulation was detected. Accordingly, even though the 16S rDNA of the bacterium was only detected in the compost soils by a nested PCR, the presence of the 16S rDNA molecules of commensal thermophile and accumulation of indole in all the enriched cultures appeared to indicate that the commensal thermophile is widely distributed in various soils.

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New Gene Cluster from Thermophile Bacillus fordii MH602 for Conversion of DL-5-Substituted Hydantoins to L-Amino Acids

  • Mei, Yan-Zhen;Wan, Yong-Min;He, Bing-Fang;Ying, Han-Jie;Ouyang, Ping-Kai
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1497-1505
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    • 2009
  • The thermophile Bacillus fordii MH602 was screened for stereospecifically hydrolyzing DL-5-substituted hydantoins to L-$\alpha$-amino acids. Since the reaction occurs at higher temperature, the advantages for enhancement of substrate solubility and for racemization of DL-5-substituted hydantoins during the conversion were achieved. The hydantoin metabolism gene cluster from thermophile is firstly reported in this paper. The genes involved in hydantoin utilization (hyu) were isolated on an 8.2-kb DNA fragment by restriction site-dependent PCR, and six ORFs were identified by DNA sequence analysis. The hyu gene cluster contained four genes with novel cluster organization characteristics: the hydantoinase gene hyuH, putative transport protein gene hyuP, hyperprotein gene hyuHP, and L-carbamoylase gene hyuC. The hyuH and hyuC genes were heterogeneously expressed in E. coli. The results indicated that hyuH and hyuC are involved in the conversion of DL-5-substituted hydantoins to an N-carbamyl intermediate that is subsequently converted to L-$\alpha$-amino acids. Hydantoinase and carbamoylase from B. fordii MH602 compared respectively with reported hydantoinase and carbamoylase showed the highest identities of 71% and 39%. The novel cluster organization characteristics and the difference of the key enzymes between thermopile B. fordii MH602 and other mesophiles were presumed to be related to the evolutionary origins of concerned metabolism.

Studies on the Tryptophanase of Thermophilic Bacteria -Part I. Screening of Indole-Forming Thermophile (고온균(高溫菌)의 tryptophanase에 관한 연구 -indole 생성균(生成菌)의 분리(分離))

  • O, Man Jin;Kim, Chan Jo
    • Korean Journal of Agricultural Science
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    • v.7 no.1
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    • pp.44-51
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    • 1980
  • The population of indole-forming thermophile was obtained for the first time several samples of soil and compost manure. The population was highly enriched by successive transfer in the tryptophan liquid medium treated with antibiotics and dilution subculture cloning. Both indole-forming thermophile might be a Gram-negative bacterium hitherto unknown, but its pure culture has not yet been established due to the inability to grow on the solid media.

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Fungal flora of paddy field in Korea IV. Filamentous fungi isolated by heat treatment (한국(韓國) 논토양중(土壤中)의 균류(菌類)에 관(關)한 연구(硏究) IV. 열처리(熱處理)로 분리(分離)한 사상균류(絲狀菌類))

  • Min, Kyung-Hee;Ito, Tadayoshi;Yokoyama, Tatsuo
    • The Korean Journal of Mycology
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    • v.15 no.3
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    • pp.187-195
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    • 1987
  • Two kinds of heat treatment method for the selective isolation of soil fungi to eliminate the commonest fungi and also to examine the vertical and seasonal distributions of the fungal population were applied to soil samples from two plots around Seoul area. The incubation method at $42^{\circ}C$ and heat treatment at $70^{\circ}C$ were used in this experiment. In the incubation method, the almost all the fungi isolated from two plots were mesophile, while the thermotolerant fungi was Aspergillus fumigatus and thermophilic fungi were Sporotrichum thermophile and Malbranchea pulchella var. sulfrea. The most dominant species isolated by this method was A. fumigatus. Nine genera and fourteen species were isolated from the two plots, and S. thermophile, Talaromyces ucrainicus,Malbranchea pulchella var. sulfrea were new to Korea. From the selection method by heat treatment at $70^{\circ}C$, ten genera and twenty species were isolated. Among these, the most fungi were also mesophile and thermotolerant fungus was A. fumigatus. The most dominant species isolated by this method was T. stipitatus, Talaromyces helicus var. major, Emericella nidulans var. nidulans, Chaetomium subspirale and Neosartorya fisheri var. fisheri were new to Korea. From the two isolation methods, it was found that the total number of soil fungi and frequency of species appeared including dominant ones were the highest at the soils of upper layer while the lowest at the soils of lower layer in its vertical distribution.

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Novel $\alpha$-Glucosidase from Extreme Thermophile Thermus caldophilus GK24

  • Nashiru, Oyekanmi;Koh, Suk-Hoon;Lee, Se-Yong;Lee, Dae-Sil
    • BMB Reports
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    • v.34 no.4
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    • pp.347-354
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    • 2001
  • $\alpha$-Glucosidase of an extreme thermophile, Thermus caldophilus GK24 (TcaAG), was purified 80-fold from cells to a homogeneous state and characterized. The enzyme exhibited optimum activity at pH 6.5 and $90^{\circ}C$, and was stable from pH 6.0 to 85 and up to $90^{\circ}C$. The enzyme had a half-life of 85 minutes at $90^{\circ}C$. An analysis of the substrate specificity showed that the enzyme hydrolyzed the non-reducing terminal unit of $\alpha$-1,6-glucosidic linkages of isomaltosaccharides and panose, $\alpha$-1,3-glycosidic bond of nigerose and turanose, and $\alpha$-1,2-glycosidic bond of sucrose. The gene encoding the TcaAG was cloned, sequenced, and sequenced in E. coli. The nucleotide sequence of the gene encoded a 530 amino acid polypeptide and had a G+C content of 68.4% with a strong bias for G or C in the third position of the codons (93.6%). A sequence analysis revealed that TcaAG belonged to the $\alpha$-amylase family. We suggest that this monomeric, thermostable, and broad-acting $\alpha$-glucosidase is a departure from previously exhibited specificities. It is, therefore, a novel $\alpha$-glucosidase.

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A New Thermophile Strain of Geobacillus thermodenitrificans Having L- Arabinose Isomerase Activity for Tagatose Production

  • Baek, Dae-Heoun;Lee, Yu-Jin;Sin, Hong-Sig;Oh, Deok-Kun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.312-316
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    • 2004
  • Five strains, producing bacterial thermostable L-arabinose isomerase, were isolated from Korean soil samples obtained from compost under high temperature circumstances. Among these strains, the CBG-Al showed the highest L-arabinose isomerase activity at $60^\circ{C}$ and was selected as a D-tagatose producing strain from D-galactose. This strain was identified as Geobacillus thermodenitrificans based on the 16S rRNA analysis, and biological and biochemical characteristics. The isolated strain was aerobic, rod-shaped, Gram-positive, nonmotile, and an endospore-forming bacterium. No growth was detected in culture temperature below $40^\circ{C}$. The maximum growth temperature and maximum temperature of enzyme activity were $75^\circ{C}$ and $65^\circ{C}$, respectively. In metal ion effects, $Ca^{2+}$ was the most effective enzyme activator with the reaction rate by 150%. In a 5-1 jar fermentor with 3-1 MY medium, L-arabinose isomerase activity was growth-associated and pH decreased rapidly after the initial logarithmic phase.

Purification and Characteristics of Adenylate Kinase from Extreme Thermophile Thermus caldophilus GK-24 (고도 호열성균 Thermus caldophilus Adenylate Kinase의 정제와 성질)

  • ;Ohta Takahisa
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.393-397
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    • 1988
  • The adenylate kinase was purified from an extreme thermophile by adenosine-pentaphospho-adenosine elution from phosphocellulose column. The molecular weight was estimated to be 22,000 by SDS-PAGE and gel filtration. The optimum temperature of the enzyme activity was 8$0^{\circ}C$ and the activation energy was given as 22.4 kcal/mole. The enzyme even showed full activity after incubation at 9$0^{\circ}C$ or in 6M guanidine-HCI at 3$0^{\circ}C$ and retained 75% of its original activity even after 1 hour at 10$0^{\circ}C$. The Michaelis constants of the enzymes for AMP, ADP, and ATP were 0.01mM, 0.017mM and 0.067mM, respectively.

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