• Korean Journal of Soil Science and Fertilizer
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• v.49 no.4
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• pp.401-408
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• 2016

The present study described the isolation of 2-methylbutyric acid (2-MBA) produced from Bacillus pumilus L1, to subsequently investigate its nematicidal activity for the control of the root-knot nematode. The results showed that 2-MBA could be purified by chromatographic techniques and was identified using nuclear magnetic resonance and liquid chromatography-mass spectrometry. Crude extract and partially purified compounds had a significant effect on the inhibition of egg hatchability and second-stage juvenile (J2) mortality. A dose-dependent effect of 2-MBA was observed for J2 mortality and egg hatchability. Egg hatchability was 69.2%, 59.9%, 32.7%, and 0.0% at 125, 250, 500, and $1000{\mu}g\;mL^{-1}$ of 2-MBA after 4 d of incubation, respectively. Meanwhile, J2 mortality was in the range of 24.4%-100.0% after 2 d of incubation, depending on the concentrations of 2-MBA used. A pot experiment also demonstrated that treatment of B. pumilus L1 culture caused a significant reduction in the number of galls, egg masses, and J2 population than that of the tap water (TW) control. However, as the B. pumilus L1 culture concentration was decreased, the efficacy of nematode control by treatment of B. pumilus L1 culture was reduced compared to that of TW. B. pumilus L1 inoculation at different concentrations also promoted cucumber plant growth. Therefore, our study demonstrated the potential of 2-MBA from B. pumilus L1 as a biocontrol agent against the root-knot nematode and a plant growth promoter for cucumber plants.

• Microbiology and Biotechnology Letters
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• v.32 no.4
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• pp.291-296
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• 2004

To produce functional and high nutritional chungkook-jang, an immuno-stimulating and rapid growing bacterium was isolated from one of Korean traditional food, chungkook-jang. The isolated bacterium was identified as Bacilpumilus and deposited in Korean Collection for Type Culture (KCTC 10461BP). The chungkook-jang fermented by JB-l has good taste and pleasant smell with high content of free amino acids, and the water extract of chungkook-jang showed 410% of immuno-stimulation activity at concentration of 2 mg/ml. The rapid fermentation in 16 h is achieved by inoculation of 20 ml culture broth of JB-l into 6 kg cooked soybean in commercial scale at $40^{\circ}C$.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.154-159
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• 2004

Bacillus species were isolated from swine manure to develope the microbial additive suitable for the rapid com-posting. The 3 of 4 isolated strains were identified as Bacillus cereus KD-2, B. pumilus KD-3, and B. licheni-formis KD-4. Bacillus sp. KD-1 was, however, not highly identical with any Bacillus sp. The isolated strains were analyzed their growth rates, enzyme activities, and antibacterial activities. The maximum growth tem-peratures of KD-1, KD-2, KD-3 and KD-4 were $45^{\circ}C$, $50^{\circ}C$, $53^{\circ}C$, and $55^{\circ}C$, respectively. The activities of pro-tease or amylase in mixed culture of 4 strains were similar in the range of $37^{\circ}C$ to $53^{\circ}C$ and activities of lipase in the range of $37^{\circ}C$ to $42^{\circ}C$ were twice higher than those of lipase in the range of $47^{\circ}C$ to $53^{\circ}C$. The antibacterial activity of KD-l, KD-2, or KD-3 against each other was not detected. That of KD-4 against KD-1, KD-2, or KD-3 was, however, detected. The organic compound and C/N ratio of compost fermented by the mixed culture were determined as 61.9% and 22.4%, respectively. The concentration of the ammonia gas was 12.35 mg/l in the liquid compost.

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.304-309
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• 2007

Soybean is useful source of protein, especially in Asia. But soybean needs heat inactivation or fermentation process before consumption, since it contains the toxic lectin and various protease inhibitors. Therefore, production of soybean boiling-waste liquor (SBWL) as a byproduct is inevitable. In this study, the chemical composition of SBWL and the optimization of culture conditions for Bacillus pumilus JB-1, a selected strain for functional chungkuk-jang fermentation, using SBWL were investigated. The SBWL contains 88% water, 9.5% free sugar, 1.6% crude protein, 0.3% crude fat, 0.1% crude fiber and 2.1% ash, respectively. The contents of total polyphenol, total flavonoids and free amino acid in SBWL were 55%, 76%, and 30% of those of raw soybean, respectively. Culture conditions for B. pumilus JB-1 in SBWL were optimized. The 1/10-diluted, 0.1 % of $(NH_4)_2SO_4$ added SBWL without pH adjustment and carbon source addition was cultured at $37^{\circ}C$ for 48 h with agitation (120 rpm). The 0.5% of inoculation was enough. The large scale fermentation in 5-L jar fermentor showed that the SBWL is a good resource for production of chungkuk-jang starter and functional ingredients.

• Food Science and Biotechnology
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• v.18 no.4
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• pp.1035-1037
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• 2009

In this study we analyzed the dynamic changes in microbiota composition during kochujang fermentation at $30^{\circ}C$. During fermentation, the viable cell counts slowly increased and reached $3.2{\times}10^7$ for aerobic bacteria, $8.3{\times}10^3$ for yeast, and $1.4{\times}10^3$ CFU/mL for fungi after 60 days. Bacilli were found to be the most dominant microorganisms throughout the fermentation process. Using the culture dependent method Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloquefaciens were found to be the main species during the early stages of fermentation; however, Bacillus pumilus and Bacillus stearothermophilus became the most dominant species during the late stage of fermentation. In contrast, when the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method was used Bacillus ehimensis was found to be the dominant species during the early stage of fermentation and Bacillus megaterium, B. pumilus, B. subtilis, and B. licheniformis were dominant in the ate stages. These results indicate various other Bacillus species rather than just B. subtilis and B. licheniformis might be involved in the fermentation of kochujang.

• KSBB Journal
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• v.26 no.5
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• pp.465-470
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• 2011

Twenty six microorganisms were isolated from soil and horse manure samples from in Iowa, U.S. Microorganisms were cultivated and screened by using plate count agar (PCA) at $35^{\circ}C$ containing 1% (w/v) oat spelt xylan instead of glucose. The xylanase activities of bacterial strains were analyzed by measuring the concentration of reducing sugar by DNS method. All isolated strains were characterized as the rod form and gram positive strains. Among the isolated strains, the HM6 strains gave the highest xylanase activity. This strain was identified as Bacillus pumilus HM6 by 16S rDNA sequence, morphological and biochemical analysis. Optimal culture temperature and initial medium pH for B. pumilus HM6 were $30-35^{\circ}C$ and pH 6-7, respectively. The maximum xylanase activity of 6879 IU/mL was obtained after growth of HM6 with 1% (w/v) oat spelt xylan at $35^{\circ}C$ for 6 days. Studies on enzymatic properties showed that the optimum conditions for the highest xylanase activity were $60^{\circ}C$ and pH 8.0. In addition, xylanase activity was stable over 2 hours at $50^{\circ}C$, whereas activity decreased after 30 min at $70^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.892-896
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• 2003

High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

• Journal of Microbiology and Biotechnology
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• v.8 no.1
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• pp.28-33
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• 1998

A gene coding for ${\beta}$-xylosidase from thermophilic xylanolytic Bacillus sp. KK-1 was cloned into Escherichia coli using plasmid pBR322. Recombinant plasmid DNAs were isloated from E. coli clones which were capable of hydrolyzing 4-methylumbelliferyl-${\beta}$-D xylopyranoside. Restriction analysis showed the DNAs to share a common insert DNA. Xylo-oligosaccharides, including xylotriose, xylotetraose, xylopentaose, and xylobiose were hydrolyzed to form xylose as an end product by cell-free extracts of the E. coli clones, confirming that the cloned gene from strain KK-1 is ${\beta}$-xylosidase gene. The ${\beta}$-xylosidase gene of strain KK-1 designated as xylB was completely sequenced. The xylB gene consisted of an open reading frame of 1,602 nucleotides encoding a polypeptide of 533 amino acid residues, and a TGA stop codon. The 3' flanking region contained one stem-loop structure which may be involved in transcriptional termination. The deduced amino acid sequence of the KK-1 ${\beta}$-xylosidase was highly homologous to the ${\beta}$-xylosidases of Bacillus subtilis and Bacillus pumilus, but it showed no similarity to a thermostable ${\beta}$-xylosidase from Bacillus stearothermophilus.

• Journal of Microbiology and Biotechnology
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• v.22 no.9
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• pp.1230-1236
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• 2012

Fish meal grades SL1 and SL2 from Sardine (Sardinella longiceps) and NJ from Pink Perch (Nemipterus japonicas) were evaluated as a sole source of carbon and nitrogen in the medium for alkaline protease production by Bacillus pumilus MTCC 7514. The analysis of the fish meal suggests that the carbon and nitrogen contents in fish meal are sufficient to justify its choice as replacement for other nutrients. Protease production increased significantly (4,914 U/ml) in medium containing only fish meal, compared with the basal medium (2,646 U/ml). However, the elimination of inorganic salts from media reduced the protease productivity. In addition, all the three grades of fish meal yielded almost the same amounts of protease when employed as the sole source of carbon and nitrogen. Nevertheless, the best results were observed in fish meal SL1 medium. Furthermore, protease production was enhanced to 6,966 U/ml and 7,047 U/ml on scaling up from flask (4,914 U/ml) to 3.7 and 20 L fermenters, respectively, using fish meal (10 g/l). Similarly, the corresponding improvement in productivities over flask (102.38 U/ml/h) was 193.5 and 195.75 U/ml/h in 3.7 and 20 L fermenters, respectively. The crude protease was found to have dehairing ability in leather processing, which is bound to have great environmental benefits.

• The Plant Pathology Journal
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• v.20 no.1
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• pp.63-66
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• 2004

Ginseng (Panax ginseng) is one of the most widely cultivated medicinal herbs in Korea. However, yield losses reached up to 30-60％ due to various diseases during 3 or 5 years of ginseng cultivation in the country. Therefore, successful production of ginseng roots depends primarily on the control of diseases. The objective of this study was to select potential biocontrol agents from rhizobacteria isolated from various plant internal root tissues for the control of multiple ginseng diseases as an alternative to fungicides. Among 106 Bacillus strains, two promising biocontrol agents, Bacillus pumilus strain B1141 and Paenibacillus lentimobus strain B1146, were selected by screening against root rot of ginseng caused by Cylindrocarpon destructans in a greenhouse. Pre-inoculation of selected isolates to seed or l-year-old root of ginseng resulted in stimulation of shoot and/or root growth of seedlings, and successfully controlled root rot caused by C. destructans (P<0.05). Furthermore, drenching of cell suspension of the selected isolates on seedling-growing pots reduced the incidence of Phytophthora blight after the seedlings were challenged with zoospores of Phytophthora cactorum (P<0.05). P. lentimorbus strain B1146 showed antifungal activity against various soil-borne pathogens in vitro, while B. pumilus strain B1141 did not show any. Results of this study suggest that some rhizobacteria can induce resistance against various plant diseases on ginseng.