Development of a RAPD-PCR method for identification of Bacillus species isolated from Cheonggukjang

Kwon, Gun-Hee; Lee, Hwang A; Park, Jae Young; Kim, Jong Sang; Lim, Jin Kyu; Park, Cheon-Seok; Kwon, Dae Young; Kim, Yong-Suk; Kim, Jeong Hwan

doi:10.1016/j.ijfoodmicro.2008.12.013

Cited by 85 publications

(72 citation statements)

References 16 publications

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“…The speciesspecific primer has been established for B. subtilis based on Endo-beta 1,4-glucanase gene and ytcP (encoding a hypothetical protein similar to a ABC-type transporter) gene (Ashe et al, 2014;Kwon et al, 2009). To the best of our knowledge, there were no such studies, in the identification of B. subtilis at subspecies level.…”

Section: Resultsmentioning

confidence: 99%

Use of novel specific primers targeted to pheS and tuf gene for species and subspecies identification and differentiation of the Bacillus subtilis subsp. subtilis, Bacillus amyloliquefaciens and Bacillus licheniformis

Huang¹,

Huang²,

Chang³

et al. 2017

Afr. J. Microbiol. Res.

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To clearly delimit the members of the Bacillus subtilis group (BSG), is difficult using common phenotypic and genotypic methods. This study described the use of pheS and tuf gene as targets for interspecies discrimination within the BSG, and also to develop specific PCR and SNP primers for species and subspecies identification and differentiation. The average sequence similarity values of the pheS and tuf gene among type strains were 85.1 and 94.7%, respectively, and all members of the BSG could be clearly distinguished based on phylogenetic analyses of pheS gene sequence. In addition, the specific primers were designed according to pheS and tuf gene sequence. The primers were shown to specifically identify B. subtilis subsp. subtilis, B. amyloliquefaciens and Bacillus licheniformis, and clearly differentiate the subspecies of B. amyloliquefaciens using specific-PCR, combined with two-plex minisequencing method. In conclusion, we have successfully established a comparative sequence analysis and rapid molecular diagnosis techniques for determination of interspecies within the BSG.

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Section: Resultsmentioning

confidence: 99%

Use of novel specific primers targeted to pheS and tuf gene for species and subspecies identification and differentiation of the Bacillus subtilis subsp. subtilis, Bacillus amyloliquefaciens and Bacillus licheniformis

Huang¹,

Huang²,

Chang³

et al. 2017

Afr. J. Microbiol. Res.

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“…Nattokinase는 serine protease이며 fibrin을 분해하는 특성을 지니고 있다 [16]. 혈전용해효소를 생 산하는 균주 중에서도 Bacillus amyloliquefaciens에 속하는 Bacillus amyloliquefaciens CH86-1 [12], Bacillus amyloliquefaciens DC-4 [15], Bacillus amyloliquefaciens DJ-4 [9] (Fig. 5).…”

Section: 서 론unclassified

Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme

Choi¹,

Lee²,

Bae³

et al. 2013

Journal of Life Science

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To isolate the fibrinolytic enzyme, 268 strains from 21 samples were morphologically isolated from Cheonggukjang collected from Korea and Japan. Among the 268 strains, protease-producing bacteria were isolated in nutrient agar medium including 1% skimmed milk. As a result of this, 22 strains were isolated. Apiweb site was used to identify these strains based on their biochemical properties. In addition, 16S rRNA sequencing was performed to identify the strain. Most of the identified strains were Bacillus subtilis and B. amyloliquefaciens. Fibrinolytic enzyme activity was measured with the fibrin plate method. Five strains were finally selected: A2-14, A2-20, C1-05, C1-09, and F2-01. Of those five strains, the A2-20 strain, which is close to B. amyloliquefaciens, showed the strongest fibrinolytic activity. The fibrinolytic enzyme produced by the A2-20 strain was partially purified from culture supernatant by gel filtration and ion exchange chromatography. The optimal pH and temperature values of the partially purified enzyme were 7.0 and 35°C, respectively. Purified protein analysis was carried out with SDS-PAGE and zymography. A genetic analysis was also conducted by PCR based on the consensus sequence of fibrinolytic enzyme. Corresponding genes with a partial sequence of the A2-20 strain were identified.Key words : Bacillus amyloliquefaciens, Cheonggukjang, enzyme purification, fibrinolytic enzyme, protease *Corresponding author *Tel：+82-51-510-2271, Fax：+82-51-514-1778 *E-mail : hsubaik@pusan.ac.kr This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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“…Bacillus amyloliquefaciens CH51 was isolated from cheonggukjang prepared at Sunchang county, Jellabukdo, Korea [9]. Bacterial strains and plasmids used are shown in Table 1.…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

“…Cloning of xynA Bacillus amyloliquefaciens CH51 was grown overnight in LB and chromosomal DNA was prepared [9]. PCR was done using a MJ mini gradient thermal cycler (BioRAD, Hercules, CA, USA).…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Cloning of a Family 11 Xylanase Gene from Bacillus amyloliquefaciens CH51 Isolated from Cheonggukjang

et al. 2012

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Development of a RAPD-PCR method for identification of Bacillus species isolated from Cheonggukjang

Cited by 85 publications

References 16 publications

Use of novel specific primers targeted to pheS and tuf gene for species and subspecies identification and differentiation of the Bacillus subtilis subsp. subtilis, Bacillus amyloliquefaciens and Bacillus licheniformis

Use of novel specific primers targeted to pheS and tuf gene for species and subspecies identification and differentiation of the Bacillus subtilis subsp. subtilis, Bacillus amyloliquefaciens and Bacillus licheniformis

Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme

Cloning of a Family 11 Xylanase Gene from Bacillus amyloliquefaciens CH51 Isolated from Cheonggukjang

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