• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.1992-1998
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• 2018

In 2015, Bacillus paralicheniformis was separated from B. licheniformis on the basis of phylogenomic and phylogenetic studies, and urease activity was reported as a phenotypic property that differentiates between the two species. Subsequently, we have found that the urease activity of B. paralicheniformis is strain-specific, and does not reliably discriminate between species, as strains having the same urease gene cluster were identified in B. licheniformis and B. sonorensis, the closest relatives of B. paralicheniformis. We developed a multilocus sequence typing scheme using eight housekeeping genes, adk, ccpA, glpF, gmk, ilvD, pur, spo0A, and tpi to clearly identify B. paralicheniformis from closely related Bacillus species and to find a molecular marker for the rapid identification of B. paralicheniformis. The scheme differentiated 33 B. paralicheniformis strains from 90 strains formerly identified as B. licheniformis. Among the eight housekeeping genes, spo0A possesses appropriate polymorphic sites for the design of a B. paralichenofomis-specific PCR primer set. The primer set designed in this study perfectly separated B. paralicheniformis from B. licheniformis and B. sonorensis.

• Biomolecules & Therapeutics
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• v.7 no.4
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• pp.385-389
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• 1999

The injibitory effect of Bacillus licheniformis AJ isolated from genitourinary normal flora as a new probiotics on the growth of diarrheal pathogens was studied. This B. licheniformis AJ inhibited the growth of E.coli O-157. Salmonella typhi and Shigella sonnei as well as the infectivity of rotavirus. However, it did not inhibit the growth of Helicobacter pyloriand human intestinal bacteria although it inhibited the harmful enzyme activity of human intestinal bacteria. B. licheniformis AJ seems to excret heat-lable growth-inhibitory protein, bacteriocin, into the media. These results suggest that B. lichenoformis AJ could be used as a new type of probiotics.

• Journal of Mushroom
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• v.14 no.4
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• pp.244-248
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• 2016

In order to isolate thermophilic bacteria with high activity of CMCase and xylanase, spent mushroom substrates was collected from an oyster mushroom cultivation farm in Jinju, Gyeongnam, Korea. Among the isolates, one strain designated as YJ09 was selected by agar diffusion method. The isolate YJ09 was identified as a member of Bacillus licheniformis based on biochemical characteristics using Bacillus ID kit and MicroLog system. Comparative 16S rDNA sequence analysis showed that isolate YJ09 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus licheniformis with sequence similarity of 98.9%. Based on its physiological properties, biochemical characteristics and phylogenetic distinctiveness, the isolate YJ09 was classified as Bacillus licheniformis. The CMCase and xylanase activity of B. licheniformis YJ09 was slightly increased corresponding to the bacterial population from exponential phase to stationary phase in the growth curve of B. licheniformis YJ09.

• Korean Journal of Veterinary Service
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• v.39 no.1
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• pp.1-6
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• 2016

In this study, Bacillus licheniformis which has been used as probiotics was isolated from Korean traditional fermented soybean. A total of 69 strains were presumptively identified as B. licheniformis by phenotypic methods. Based on PCR amplification and 16S rRNA gene sequencing, the multilocus sequence typing of gyrA and rpoB, followed by phylogenetic analysis was performed. The isolates were distinctly differentiated and found to be closely related to B. amyloliquefaciens, B. subtilis, and B. aerius. The partial 16S rRNA gene sequences of those strains matched those of B. sonorensis (97%) and B. aerius (98%) in the phylogenetic tree. In contrast, multilocus phylogenetic analysis (MLPA) showed that only 61 (86.9%) out of 69 strains were B. licheniformis. The rest of those strains were found to be B. subtilis (5.8%), B. amyloliquefaciens (2.9%), and B. sonorensis (2.9%), respectively. Therefore, our results suggested that since the 16S rRNA gene sequencing alone was not sufficient to compare and discriminate closely related lineages of Bacillus spp., it was required to analyze the MLPA simultaneously to avoid any misleading phenotype-based grouping of these closely related species.

• The Korean Journal of Mycology
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• v.49 no.2
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• pp.199-209
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• 2021

The present study was performed to improve the technique used for fermenting the mushroom growth medium. Taxonomic analysis of 16S rDNA sequence from the predominant Bacillus strain CY-24 isolated during the fermentation phase of the rice straw medium identified it as Bacillus licheniformis. In addition, the growth environment of B. licheniformis was also examined in this study, which revealed the optimal growth temperature and pH to be 30 ℃ and 6.0, respectively. This study also revealed that carboxymethyl cellulase (CMCase) and polygalacturonase (PGase) enzymes isolated from B. licheniformis achieved their maximal activities at 50 ℃ and 60 ℃ respectively. Furthermore, the study confirmed that the two enzymes, i.e., CMCase and PGase in B. licheniformis are stable at temperatures above 60 ℃. The present study thus demonstrates that B. licheniformis CY-24 possesses excellent enzymatic properties. It also reveals that the action of enzymes during the production of growth mediums used for the cultivation of mushrooms is closely associated with the promotion of fermentation and softening of the rice straw. Overall, this study provides elementary information regarding the role of B. licheniformis enzymes during growth medium fermentation for Agaricus bisporus cultivation.

• Applied Biological Chemistry
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• v.40 no.3
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• pp.173-177
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• 1997

Bacillus licheniformis 9945a releases a natural ${\gamma}-poly(glutamic\;acid)({\gamma}-PGA)$ into fermentation broth and shows a mucoid phenotype on the solid agar medium. Transformation of mucoid cells of Bacillus species has not been simple and straightforward. The transpositional activity of Tn10 in B. licheniformis also has not been own either. Thus, a spontaneous non-mucoid derivative of the B. licheniformis was obtained first. Shuttle vector pHV1248 containing mini-Tn10 was introduced into the non-mucoid derivative by the method of protoplast transformation. The resulting transformant was reverted to the wild mucoid phenotype, and then mutated randomly with the mini-transposon by heat induction. Auxotrophs requiring arginine, lysine, or tryptophan were isolated by replica plating method. Southern blotting and DNA-DNA hybridzation analysis showed that these auxotrophs were generated by mini-Tn10 insertion into the chromosomal DNA. This method of transformation and mutation using pHV1248 would be useful for the generation of diverse mutants of B. licheniformis 9945a.(Received January 24,1997; accepted March 10, 1997)

• Journal of Life Science
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• v.17 no.7 s.87
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• pp.983-989
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• 2007

Bacillus lichemiformis Kll, a plant growth promoting rhizobacterium was reported as a producer of auxin, siderophore, as well as antifungal cellulase under some culture conditions. In vitro test, B. licheniformis Kll represented excellent antagonistic ability against Fusarium oxyspoum (KACC 40037), and showed broad spectrum against other phytopathogenic fungi. B. licheniformis Kll had cellulolytic activity toward not only carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as fungal cell wall cellulose, filter paper (Whatman No. 1), and Avicel. In addition, we confirmed antifungal substance production by butanol-extract methods. The strain produced optimally the antifungal substance when it was cultivated at pH 9.0, 30${\circ}$C for 4 days on nutrient medium. The biological control mechanisms of B. lichemiformis Kll were caused by antifungal substance, cellulase and siderophore against phytopathogenic fungi.

• Journal of Applied Biological Chemistry
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• v.52 no.3
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• pp.121-125
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• 2009

We measured the influence of antifungal antagonists Bacillus subtilis AH18 and Bacillus licheniformis K11 on soil microbial community in microcosms. Both antifungal antagonists were confirmed to suppress hot pepper phytophthora blight. Phospholipid fatty acids (PLFA) were analyzed to investigate the soil microbial community. B. subtilis AH18 changed the total PLFA composition and bio-indicators of PLFA, compared with other treatments. B. subtilis AH18 decreased the proportion of bacteria and gram negative/gram positive bacteria, and increased the fungi/bacteria and anaerobic/aerobic microorganisms. In addition cy19:0/18:$1{\omega}7c$, which means adaptation to unfavorable environmental conditions, was increased by the application of B. subtilis AH18. On the other hand the inoculation of B. licheniformis K11 or combined inoculation of both antifungal strains did not affect soil microbial community. The suppression of phytophthora blight and preservation of indigenous soil microbial community may be achieved by the combined application of B. subtilis AH18 and B. licheniformis K11.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.3
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• pp.526-533
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• 1999

Two species of antimicrobial agent producing bacteria and one sensitive strain were isolated from soil. Those were identified as B. subtilis, B. licheniformis, and Curtobacterium sp. by morphological, biochemical, physiological and chemotaxonomic characteristics. These were designated as B. subtilis cx1, B. licheniformis cy2 and Curtobacterium sp. cf3, respectively. Antimicrobial agent produced by B. subtilis cx1 showed high antibacterial activity against gram positive bacteria including of B. subtilis, Curtobacterium sp., L. mesenteroids, Staphy. aureus, S. faecalis and even gram negative bacteria, P. aeruginosa. Antimicrobial agent from B. licheniformis cy2 showed slightly lower antimi crobial activity than that from B. subtilis cx1. These two strains showed maximum production of antimicrobial agents at 30oC for 9~21hr cultivation. Curtobacterium sp. cf3 showed more sensitive activity than a sensitive strain of B. subtilis ATCC 6633 which was same genus or species with the B. subtilis cx1 and B. subtilis cy2, when the antimicrobial agent producing strains, B. subtilis cx1 and B. subtilis cy2, were directly applied onto these sensitive lawns.

• Korean Journal of Environmental Agriculture
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• v.22 no.3
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• pp.203-209
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• 2003

In the course of screening for antifungal agents, a bacterial strain, DM3 was isolated from a mud sample collected at Daechon in Chungnam province and identified as Bacillus licheniformis based on API 50CHB test. It has antifungal activity against 12 plant pathogenic fungi in paper disc assay. At the 95% lethal dose of gamma radiation ($^{60}Co$, 10 kGy, $D_{10}=2.32\;kGy$), the antifungal activity deficient mutant (mDM3) against Colletotrichum gloeosporioides was induced From 2-D electrophoresis analysis, serine hydroxymethyltransferase (45.0 kDa), hypothetical protein(40.7 kDa), NifU protein homolog(15.4 kDa), and resolvase(12.5 kDa) homologous proteins were detected only in B. licheniformis DM3. Lysozyme(18.1 kDa) and alkyl hydroperoxide reductase(15.6 kDa) homologous proteins were expressed uniquely in B. licheniformis mDM3. Further studies are needed to reveal that these proteins from B. licheniformis DM3 could be closely related to the antifungal activity against plant pathogenic fungi.