경남지역의 큰느타리버섯재배사에서 채취한 버섯폐배지로부터 그람양성이며 내생포자를 형성하는 간균을 분리하여 이를 CS9라 명명하였다. 분리균 CS9는 통성혐기성균으로 운동성이 있으며 xylanase를 생성하였다. 분리균의 최적 생육조건은 $40^{\circ}C$, pH 6.0이었고, 지방산 조성분석 결과 세포벽을 이루는 주요지방산은 anteiso-C15:0, anteiso-C17:0, iso-C17:0이었으며, DNA G+C함량은 45 mol%로 Bacillus 속의 G+C함량인 43-46 mol%의 범위에 속하는 것으로 확인되었다. 분리균의 16S rDNA 염기서열을 분석한 결과, 분리균 CS9은 Bacillus subtilis YB1 균주와 96.8%의 유사도를 나타내었고, Bacillus속의 다른 균주들과는 92-94%의 유사도를 나타내었다. 이상의 생리적, 생화학적, 유전적 특성을 근거로 큰느타리버섯폐배지로부터 분리한 xylanase를 생성하는 CS9는 Bacillus subtilis CS9로 동정되었다.
경남지역의 큰느타리버섯재배사에서 채취한 버섯폐배지로부터 그람양성이며 내생포자를 형성하는 간균을 분리하여 이를 CS9라 명명하였다. 분리균 CS9는 통성혐기성균으로 운동성이 있으며 xylanase를 생성하였다. 분리균의 최적 생육조건은 $40^{\circ}C$, pH 6.0이었고, 지방산 조성분석 결과 세포벽을 이루는 주요지방산은 anteiso-C15:0, anteiso-C17:0, iso-C17:0이었으며, DNA G+C함량은 45 mol%로 Bacillus 속의 G+C함량인 43-46 mol%의 범위에 속하는 것으로 확인되었다. 분리균의 16S rDNA 염기서열을 분석한 결과, 분리균 CS9은 Bacillus subtilis YB1 균주와 96.8%의 유사도를 나타내었고, Bacillus속의 다른 균주들과는 92-94%의 유사도를 나타내었다. 이상의 생리적, 생화학적, 유전적 특성을 근거로 큰느타리버섯폐배지로부터 분리한 xylanase를 생성하는 CS9는 Bacillus subtilis CS9로 동정되었다.
A Gram-positive, endospore-forming, rod-shaped bacterial strain was isolated from spent mushroom (Pleurotus eryngii) substrates taken from the Hoamfarm located in Keyollgnam, Korea. The isolate, designated CS9, was facultatively anaerobic, motile rod and produced xylanase. The strain grew optimally ...
A Gram-positive, endospore-forming, rod-shaped bacterial strain was isolated from spent mushroom (Pleurotus eryngii) substrates taken from the Hoamfarm located in Keyollgnam, Korea. The isolate, designated CS9, was facultatively anaerobic, motile rod and produced xylanase. The strain grew optimally at $40^{\circ}C$ and pH 6.0. The major cellular fatty acids were anteiso-$C_{15:0}$, anteiso-$C_{17:0}$, and iso-$C_{17:0}$. The genomic DNA G+C content was 45 mol%. Comparative 16S-rDNA sequence analysis showed that the isolate CS9 formed a distinct phyletic line within the genus Bacillus and was most closely related to Bacillus subtilis YB1, with 16S DNA sequence similarity of 96.8%. Sequence similarities to other type strains were 92-94%. On the based of physiological and molecular properties, the isolate CS9 was classified within the genus Bacillus as Bacillus subtilis CS9.
A Gram-positive, endospore-forming, rod-shaped bacterial strain was isolated from spent mushroom (Pleurotus eryngii) substrates taken from the Hoamfarm located in Keyollgnam, Korea. The isolate, designated CS9, was facultatively anaerobic, motile rod and produced xylanase. The strain grew optimally at $40^{\circ}C$ and pH 6.0. The major cellular fatty acids were anteiso-$C_{15:0}$, anteiso-$C_{17:0}$, and iso-$C_{17:0}$. The genomic DNA G+C content was 45 mol%. Comparative 16S-rDNA sequence analysis showed that the isolate CS9 formed a distinct phyletic line within the genus Bacillus and was most closely related to Bacillus subtilis YB1, with 16S DNA sequence similarity of 96.8%. Sequence similarities to other type strains were 92-94%. On the based of physiological and molecular properties, the isolate CS9 was classified within the genus Bacillus as Bacillus subtilis CS9.
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제안 방법
PCR reaction was typically performed in a final volumn of 100 ul using 80 pmol of each primer, 50 ng of template, Taq polymerase (Takara), and 1 mM of each dNTP. Reactions comprised 30 cycles of 94C for 1 min, annealing at 56oC for 30 sec and extention at 72oC for 1 min with a final extention at 72oC for 5 min using primers, which designed based on conserved region between 16S rDNA sequences of Bacillus species. The forward primer was 5' -TTCTACGGAGAGTTTGATCC-3' and the reverse primer was 5' —CACCTTCCGGTACGGCTACC-3'.
The strips were incubated at 40℃ for 48 h. Tests for utilization of substrates as sole carbon source were performed with BIOLOG automatic identification system (Biolog, Inc., Hayward, CaHf.). Strain was grown on complex medium at 40 ℃ for 24 h and suspended in a sterile solution containing 10% salts.
대상 데이터
Nine bacterial strains were isolated from spent mushroom {Plewotus eryngii) substrates obtained from the Hoamfarm located in Keyongnam, Korea (Fig. 1). From nine strains, one positive strain surrounded by the clear ring that indicates xylanase activity, designated CS9, was isolated on LB agar (Difco) supplemented with 0.
이론/모형
by saponification. Fatty acid methyl esters were prepared using the method described in the MIDI Microbial Identification System (Hewlett Packard) manual. The resulting esters were separated by gas chromatography (model 5890, Hewlett Packard) fitted with a phenylmethyl silicone fused silica capillary column (25 m, 0.
Amplified 16S rDNA fragments were used as sequencing templates. Nucleotide sequencing was done by the dideoxy-chain termination method of Sanger et al. (1977).
Canada). Phylogenetic comparison was inferred by using the neighbour-joining method. The resulting tree was drawn with the DNAMan software and treated as rooted, although the outgroup designation option was included to polarize character states.
성능/효과
4) constructed from the sequence data shows that strain CS9appeared within the evolutionary radiation encompassing the genus Bacillus and occupied a distinct phylogenetic position within the genus. The 16S rDNA sequence of isolate CS9 was most closely related to Bacillus subtilis YB1, with 16S DNA sequence similarity of 96.8%. Levels of 16S rDNA similarity between strain CS9 and other Bacillus species were 92—94%.
0이었고, 지방산 조성분석 결과 세포벽을 이루는 주요지방산은 anteiso-C15:0, anteiso—C17:0, iso—C17:0이 었으며 , DNA G+C함량은 45 mol%로 Bacillus 속의 G+C함량인 43-46 mol%의 범위에 속하는 것으로 확인되었다. 분리균의 16S rDNA 염기서열을 분석한 결과, 분리 균 CS9은 Bacillus subtilis YB1 균주와 96.8%의 유사도를 나타내었고, c/s속의 다른 균주들과는 92-94%의 유사도를 나타내었다. 이상의 생리적, 생화학적, 유전적 특성을근거로 큰느타리 버섯폐 배지로부터 분리한 xylanase를 생성하는 CS9는 Bacillus subtilis CS9로 동정 되었다.
분리균 CS9는 통성혐기성균으로 운동성이 있으며 xylanase를 생성하였다. 분리균의 최적생육조건은 40℃, pH 6.0이었고, 지방산 조성분석 결과 세포벽을 이루는 주요지방산은 anteiso-C15:0, anteiso—C17:0, iso—C17:0이 었으며 , DNA G+C함량은 45 mol%로 Bacillus 속의 G+C함량인 43-46 mol%의 범위에 속하는 것으로 확인되었다. 분리균의 16S rDNA 염기서열을 분석한 결과, 분리 균 CS9은 Bacillus subtilis YB1 균주와 96.
8%의 유사도를 나타내었고, c/s속의 다른 균주들과는 92-94%의 유사도를 나타내었다. 이상의 생리적, 생화학적, 유전적 특성을근거로 큰느타리 버섯폐 배지로부터 분리한 xylanase를 생성하는 CS9는 Bacillus subtilis CS9로 동정 되었다.
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