해안가 토양으로부터 강력한 chitinase 활성을 가진 Bacillus cereus KJA-118이 분리.동정되었다. B. cereus KJA-118은 1% colloidal chitin이 포함된 배지를 pH 6.0으로 조절한 후 $30^{\circ}C$ 에서 4일동안 호기적으로 배양했을 때 가장 높은 chitinase 효소활성을 보였다. 탄소원 (crab shell powder, chitin powder, colloidal chitin, and R. solani 균사)에 따른 chitinase 활성은 R. solani 균사를 사용했을 때 가장 높았다. Glycol chitin 0.01%가 포함된 gel에서 전기영동 후 활성 염색한 결과, B. cereus KJA-118에 의해 생산되는 chitinase는 분자량이 68, 47, 37 KDa인 3개의 isoform이 검출되었다. 액체 배지에서 미리 배양한 B. cereus KJA-118과 R. solani를 다시 혼합 배양했을 때, 곰팡이의 세포벽이 완전히 파괴되었다. R. solan가 감염된 토양에 B. cereus KJA-118의 배양액을 처리했을 때 28.1%의 오이 모잘록병 억제 효과를 확인하였다.
해안가 토양으로부터 강력한 chitinase 활성을 가진 Bacillus cereus KJA-118이 분리.동정되었다. B. cereus KJA-118은 1% colloidal chitin이 포함된 배지를 pH 6.0으로 조절한 후 $30^{\circ}C$ 에서 4일동안 호기적으로 배양했을 때 가장 높은 chitinase 효소활성을 보였다. 탄소원 (crab shell powder, chitin powder, colloidal chitin, and R. solani 균사)에 따른 chitinase 활성은 R. solani 균사를 사용했을 때 가장 높았다. Glycol chitin 0.01%가 포함된 gel에서 전기영동 후 활성 염색한 결과, B. cereus KJA-118에 의해 생산되는 chitinase는 분자량이 68, 47, 37 KDa인 3개의 isoform이 검출되었다. 액체 배지에서 미리 배양한 B. cereus KJA-118과 R. solani를 다시 혼합 배양했을 때, 곰팡이의 세포벽이 완전히 파괴되었다. R. solan가 감염된 토양에 B. cereus KJA-118의 배양액을 처리했을 때 28.1%의 오이 모잘록병 억제 효과를 확인하였다.
A bacterium, KJA-118 showing a strong chitinase activity, was isolated and identified as Bacillus cereus. The strain produced maximum level of chitinase, when grown aerobically at $30^{\circ}C$ for 4 days in basal broth containing 1% colloidal chitin in the initial pH adjusted to 6.0. Amo...
A bacterium, KJA-118 showing a strong chitinase activity, was isolated and identified as Bacillus cereus. The strain produced maximum level of chitinase, when grown aerobically at $30^{\circ}C$ for 4 days in basal broth containing 1% colloidal chitin in the initial pH adjusted to 6.0. Among various carbon sources such as crab shell powder, chitin powder, colloidal chitin, and R. solani mycelium, maximum chitinase activity was found in culture broth supplemented with R. solani mycelium. When KJA-118 was incubated with R. solani, the cell wall of the fungus was found to be completely destroyed. SDS-PAGE and active staining results revealed that KJA-118 produced three isoforms of chitinase with molecular weights of 68 kDa, 47 kDa, and 37 kDa. When the suspension of KJA-118 was treated to cucumber seedlings, reducing rate of damping-off caused by R. solani was about 28.1%.
A bacterium, KJA-118 showing a strong chitinase activity, was isolated and identified as Bacillus cereus. The strain produced maximum level of chitinase, when grown aerobically at $30^{\circ}C$ for 4 days in basal broth containing 1% colloidal chitin in the initial pH adjusted to 6.0. Among various carbon sources such as crab shell powder, chitin powder, colloidal chitin, and R. solani mycelium, maximum chitinase activity was found in culture broth supplemented with R. solani mycelium. When KJA-118 was incubated with R. solani, the cell wall of the fungus was found to be completely destroyed. SDS-PAGE and active staining results revealed that KJA-118 produced three isoforms of chitinase with molecular weights of 68 kDa, 47 kDa, and 37 kDa. When the suspension of KJA-118 was treated to cucumber seedlings, reducing rate of damping-off caused by R. solani was about 28.1%.
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제안 방법
The objectives of this study were to evaluate the chitinase production, antifungal activity against pathogenic fungi, and biological control against Rhizoctonia solani damping-off of cucumber with Bacillus cereus KJA-118 isolated from coastal area containing shellfish waste.
To determine isoforms and the molecular weight of chitinase produced by B. cereus KJA-118, the crude enzyme collected from culture broth containing 1% colloidal chitin grown at 3*0 0 for 4 days was analyzed using SDS-PAGE and the active staining method. As shown in Fig.
대상 데이터
The material was dehydrated with ethanol at 4℃ using a series of steps for 10 min each. The specimens were dried with CO2 as the carrier gas. The dried specimens were mounted on stubs with Television Tube Koat to prevent charging.
The soil samples obtained from a coastal area of Youngkwang at Chonnam Province, Korea were incubated in a basal medium containing 1 % colloidal chitin as a sole carbon. The composition of the basal medium was 0.
성능/효과
Partial sequence of the 16S rRNA gene of KJA-118 amplified by PCR was compared to the corresponding sequences of other bacterial 16S rRNA gene using BLAST under NCBI. Within 347 base pairs of the DNA compared, 345 bases correspond with B. cereus 16S rRNA sequences (Accession No. AF176322) where base pairs at 18 and 20 positions of B. cereus were t and g, while those of KJA-118 are c and a, respectively (data not shown), resulting in 99.4% of homology. Therefore, the bacterium was named as B.
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