• Korean Journal of Microbiology
• /
• v.42 no.4
• /
• pp.313-318
• /
• 2006

A thermophilic bacterium producing the extracellular cellulase was isolated from soybean paste, and the isolate WL-12 has been identified as Bacillus licheniformis on the basis on its 16S rRNA sequence, morphology and biochemical properties. A gene encoding the cellulase of B. licheniformis WL-12 was cloned and its nucleotide sequence was determined. This cellulase gene, designated celA, consisted of 1,551 nucleotides, encoding a polypeptide of 517 amino acid residues. The gene product contained catalytic domain and cellulose binding domain. The deduced amino acid sequence was highly homologous to those of cellulases of B. licheniformis, B. subtilis and B. amytoliquefaciens belonging to the glycosyl hydrolase family 5. When the celA gene was highly expressed using a strong B. subtilis promoter, the extracellular cellulase was produced up to 7.0 units/ml in B. subtilis WB700.

• KSBB Journal
• /
• v.23 no.3
• /
• pp.251-256
• /
• 2008

The aim of this work was to investigate the fibrinolytic activity and characterization of Bacillus licheniformis HK-12, which produces the fibrinolytic enzyme excreted from naturally fermented Chungkook-Jang. Initially, the physiological and biochemical characteristics of strain HK-12 was examined. Both physiological analysis using BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were performed to identify the strain, and the strain could be assigned to Bacillus licheniformis, designated as B. lichenformis HK-12, and registered in GenBank as [EU288193]. Phylogenetic analysis of B. licheniformis HK-12 was plotted based on 16S rRNA sequence comparisons. During the incubation period of B. licheniformis HK-12, the changes of bacterial growth, fibrinolytic activity, and pH were monitored. As the results, after 36 hours of incubation, the maximum fibinolytic activity was about 2.25 times than that of plasmin used as standard. Optimal conditions on the growth of B. licheniformis HK-12 associated with the fibrinolytic activity was initial pH 7.0 and 40$^{\circ}C$, respectively.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.31 no.2
• /
• pp.204-210
• /
• 2002

The quality characteristics and sensory evaluation of chungkookjang were investigated. The samples were prepared and fermented by the inoculation of Bacillus strains; B. subtilis, B. natto and B. licheniformis as a single starter, and mixed culture of B. natto and B. licheniformis on the industrialized model system. It was shown that microbial growth, protease activity, contents of amino-and ammonia-nitrogen and contents of organic acid were higher in B. subtilis inoculated sample, and were lower in B. licheniformis inoculated one. General quality characteristics of sample inoculated by mixed culture of B. natto and B. licheniformis took a middle position between each B. natto and B. licheniformis inoculated one. Fifty eight species of odor components were identified. Ethanol, 3-methyl-1-butanol, acetic acid, benzaldehyde and alkyl pyrazines were identified in all samples and most of other flavor components were strain specific. The contents of unpleasant smell components, alkyl pyrazines and benzaldehyde, were lower in B. licheniformis inoculated sample. The sensory evaluations showed that chungkookjang manufactured from mixed culture of B. natto and B.licheniformis was most acceptable. Therefore, results indicated that chungkookjang manufactured from mixed culture of B. natto and B. licheniformis induced better sensory quality than that of the control.

• Korean Journal of Food Science and Technology
• /
• v.17 no.1
• /
• pp.8-14
• /
• 1985

In order to give the unique flavour of traditional Doenjang (Korean-style soybean paste) to commercially manufactured soybean paste, the addition of Bacillus licheniformis and Saccharomyces rouxii was tried and their influences were investigated. The addition of B. licheniformis and S. rouxii in soybean paste decreased the content of reducing sugars. By the addition of B. licheniformis, the contents of amino type nitrogen and titrable acidity were increased. The content of ammoniacal nitrogen was reduced and ethyl alcohol production was increased by the addition of S. rouxii. The viable counts of molds showed the decreasing tendency during the aging and it was accelerated by the addition of B. licheniformis and S. rouxii. Acetaldehyde, acetone, ethylacetate, methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol and isopropyl alcohol were detected from the vapor of all tested ripen soybean pastes, while in a traditional Doenjang, isoamyl alcohol and isobutyl alcohol were not. The B. licheniformis and S. rouxii added sample showed richest free amino acid content. In organoleptic test the B. licheniformis and S. rouxii added sample showed the most excellent overall acceptability.

• Korean Journal of Microbiology
• /
• v.35 no.4
• /
• pp.296-301
• /
• 1999

A Bacillus strain from Korean soil was isolated and identified to be Bacillus licheniformis B1 through various biochemical tests, VITEK, and MIDI system analysis. The strain produced extracellular amylase and protease. Whether or not the strain can perform Chungkookjang fermentation with autoclaved soybean and Kanjang fermentation was determined in this study. In Chungkookjang fermentation, browining materials of strong anti-oxidant increased 8-fold, and 2-fold in Kanjang, compared with initiation material for fermentation. Maximal protease activity in Chungkookjang was observed one day after inoculation. Protease activities in Kanjang decreased to the half, and then maintained constant values during fermentation, probably due to the inhibitory effect of salt on protease activities. High molecular mass of nucleic acids was identified in Chungkookjang and Kanjang. Since the nucleic acids were not observed in autoclaved soybean, they seem to be originated from B. licheniformis B1. This study demonstrated successive fermentation of Chungkookjang and Kanjang by B. licheniformis B1 isolated from nature, and suggested possible development of food rich in browing and nucleic acids.

• Journal of Korean Society of Forest Science
• /
• v.104 no.3
• /
• pp.512-518
• /
• 2015

Pine wilt disease (PWD) caused by pine wood nematode, Bursaphelenchus xylophilus, has become the most serious threat to pine trees in Korea. This study was subjected to investigate effective biological control agent against PWD. To select nematocidal bacteria against PWD, Bacillus licheniformis MH48 was selected among five bacteria due to its high nematocidal potential. B. licheniformis MH48 was tested for cell growth and protease activity to evaluate its nematicidal potential. In the B. licheniformis MH48, cell numbers were highest three days after incubation, while protease activity was highest after seven days. In the effect of different concentrations of B. licheniformis MH48 culture broth against B. xylophilus, 20% concentration of culture broth showed approximately 80% of pine wood nematode mortality compared to the control. Especially, pine wood nematode's cuticle layers were degraded two days after treatment of B. licheniformis MH48 culture broth. The present study suggests that B. licheniformis MH48 can be one of the potential biocontrol candidates against pine wood nematode due to its ability to produce protease.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.30 no.2
• /
• pp.179-190
• /
• 2010

The experiment was conducted to observe the effects of dried whole crop barley treated with cellulolytic microorganisms (Aspergillus niger KCCM 60357 and Bacillus licheniformis KCCM 40934) on the chemical composition, in vitro colonic fermentation and whole tract digestibility in swine. Whole crop barley were fermented with no microorganism addition (control), A. niger, B. licheniformis and co-culture of A. niger and B. licheniformis (Mixture) for 3 days at $30^{\circ}C$. In the feed chemical composition, CP contents of whole crop barley treated with A. niger (7.52%) and B. licheniformis (7.77%) were significantly higher than control (6.81%) (p<0.05). The in vitro colonic fermentation of dried whole crop barley fermented with control showed significantly higher $CH_4$ contents than A. niger, B. licheniformis and Mixture at 18h incubation (p<0.05). Dry matter (DM) digestibilities of A. niger (55%) and Mixture (57.42%) treatments were significantly higher than control (43.74%) (p<0.05). Ammonia-N was significantly increased in A. niger, B. licheniformis and Mixture relative to control at 24 hour incubation (p<0.05). Xylanase activities in A. niger, B. licheniformis and Mixture treatments were significantly higher than control at 24 hour incubation (p<0.05). Concentrations of total VFA were significantly increased in B. licheniformis (12.61 mM) at 24hour incubation (p<0.05). In vitro whole tract digestibility was significantly increased in B. licheniformis (49.61%) compared with the control (45.65%) (p<0.05). In conclusion, whole crop barley treated with cellulolytic microorganisms improved whole tract digestibility and colonic fermentation for swine.

• Korean Journal of Food Science and Technology
• /
• v.38 no.4
• /
• pp.548-553
• /
• 2006

The aim of this study was to produce Chungkookjang-a food produced through fermentation with Bacillus licheniformis E1-that contains an increased concentration of a bacterial biological response modifier (B-BRM). Unfortunately, sensory studies have indicated that B. licheniformis E1-fermented Chungkookjang is unacceptable for commercial use. We isolated another bacterial strain from this food product: B. subtilis S2. The optimum time and temperature for Chungkookjang fermentation with B. licheniformis E1 and B. subtilis S2 were 48 hr and $40^{\circ}C$, respectively. Sensory studies showed that Chungkookjang fermented by both B. licheniformis E1 and B. subtilis S2 was more acceptable than B. licheniformis E1 only. The amino nitrogen and crude protein content of the product were 359 mg% and 45.6% respectively. Additionally, it was confirmed that the proliferation of mouse splenic lymphocytes increased significantly, when the cells were treated with the BRM from Chungkookjang fermented using the mixture of bacterial strains in vitro. These results suggest that the enriched Chungkookjang may help patients who are medically in need of potentiation of lymphocytes proliferation.

• KSBB Journal
• /
• v.29 no.3
• /
• pp.131-138
• /
• 2014

In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.

• Microbiology and Biotechnology Letters
• /
• v.40 no.4
• /
• pp.296-302
• /
• 2012

A bacterial strain was isolated from soybean paste fermented in a Korean Buddhist temple as a producer of the extracellular thermostable ${\alpha}$-amylase. The isolate YB-1234 has been identified as Bacillus licheniformis on the basis of its 16S rDNA sequence, morphology and biochemical properties. A gene encoding the thermostable ${\alpha}$-amylase of B. licheniformis YB-1234 was cloned into Escherichia coli and its nucleotide sequence was determined. The deduced amino acid sequence of ${\alpha}$-amylase was very highly homologous to those of the thermostable ${\alpha}$-amylases of B. licheniformis belonging to the glycosyl hydrolase family 13. The ${\alpha}$-amylase produced by recombinant E. coli carrying the ${\alpha}$-amylase gene exhibited maximal activity at pH 6.0, identical to ${\alpha}$-amylase in the culture filtrate of B. licheniformis, while the temperature profile was somewhat different between the two. Particularly, ${\alpha}$-amylase produced from B. lcheniformis is much more thermostable than that from recombinant E. coli. The predominant products resulting from the ${\alpha}$-amylase hydrolysis were glucose, maltose and maltotriose for maltotetraose and maltohexaose.