• Journal of Life Science
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• v.14 no.3
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• pp.391-395
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• 2004

To overproduce the cyclodextrin glucanotransferase(CGTase) of Bacillus stearothermophilus NO2 in B. subtilis, the pJH-CGTl plasmid (8.14 kb) was constructed, in which the ORF of CGTase gene could be transcribed by strong constitutive promoter, P$\_$JH/. To overproduce CGTase from a recombinant B. subtilis, the effect of media on the cell growth and expression level of CGTase were investigated with various media (LB, 2${\times}$LB, 5% molasses+2% CSL, CS, LBG) in the flask culture. Among them, [5% molasses+2% CSL] medium revealed the maximum expression level of CGTase with 1.8 unit/$m\ell$ at 9 hr culture. In the batch culture on [10% molasses+5% corn steep liquor] medium the expression level of CGTase, the secretion efficiency, and plasmid stability were about 4.2 unit/$m\ell$, 90% and 90%, respectively, at 30 hr culture. The cell growth and expression level in the fermenter culture with the industrial molasses medium were increased by 2-folds over the flask culture.

• Microbiology and Biotechnology Letters
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• v.41 no.2
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• pp.160-167
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• 2013

In order to investigate changes in quality and enzyme activity during Chungkookjang fermentation, germinated- and nongerminated yellow soybeans were fermented by Bacillus subtilis and traditional methods. When the soybean was soaked for 6 h and then watered for 4 days with 2 h-interval at $25^{\circ}C$, the highest germination rate was obtained. The germinated soybeans had a higher total isoflavone ($988.4{\mu}g/g$) than that of the nongerminated soybeans ($859.5{\mu}g/g$). Amino type nitrogen contents, protease and amylase activities were higher in germinated soybean Chungkookjang, which was fermented with B. subtilis, than nongerminated soybean Chungkookjang, which was fermented with B. subtilis and traditional methods. Reducing sugar and amino type nitrogen contents, the number of viable cells and protease and amylase activities, were higher for Chungkookjang fermented with B. subtilis, than Chungkookjang fermented by traditional methods. ALP and SOD activities in the Chungkookjang diet group were considerably higher than in the control group. AST activity in the germinated soybean Chungkookjang diet group was higher than in the nongerminated soybean Chungkookjang diet group. In conclusion, it is suggested that Chungkookjang prepared with germinated soybeans using B. subtilis D7 could be practically used as a functional product.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.307-311
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• 2005

Marine microorganisms to produce functional biopolymers were isolated from the seashore of the Kyungsang province. Microorganisms to hydrolyze carboxy-methyl cellulose(CMC) were cultured in marin broth and the other liquid medium that contained 2.0% (w/v) glucose, 0.25% yeast extract, 0.5% $K_2HPO_4$, 1% NaCl, 0.02% $MgSO_4{\cdot}7H_2O$ and 0.06% $(NH_4)_2SO_4$ to investigate the ability to produce carboxymethyl cellululase (CMCase) under aerobic conditions. Twelve microorganisms among them showed higher activities of CMCase than B. amyloliquefaciens DL-3, which was known as a cellulase-producing strain. The microorganism showing highest activity of CMCase in this study was identified as Bacillus subtilis subsp. subtilis with 16S rDNA partial sequencing and gyrase A partial sequencing and named as B. subtilis subsp. subtilis A-53.

• Journal of Life Science
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• v.23 no.1
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• pp.102-109
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• 2013

To isolate GABA-producing microorganisms, 1,500 strains were isolated from different Chungkookjang samples and screened. From these strains, 20 were selected for further analyses based on a protease and slime-producing activity test. The MC 31 strain showed the highest GABA concentration in Chungkookjang and was used in this study. MC 31 was identified as Bacillus subtilis by an API 50CHB kit and 16S rDNA sequences analysis and named as B. subtilis MC 31. B. subtilis MC 31 showed exponential growth up to 12 hours at $37^{\circ}C$ in LB broth, and it reached a stationary phase after 24 to 36 hours of incubation. B. subtilis MC 31 showed maximum GABA content at 72 hours after incubation at $40^{\circ}C$.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.207-212
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• 2010

A bacterium producing the extracellular mannanase was isolated from Korean soybean paste. The isolate WL-8 has been identified as Bacillus subtilis on the basis on its 16S rRNA sequence, morphology and biochemical properties. The mannanase productivity of strain WL-8 was increased in LB broth by addition of wheat bran. The maximum mannanase productivity was reached to approximately 20 U/ml in LB medium supplemented with 6% wheat bran. A gene encoding the mannanase of WL-8 was cloned into Escherichia coli and its nucleotide sequence was subsequently determined. The mannanase gene consisted of 1,086 nucleotides encoding a polypeptide of 362 amino acid residues. The deduced amino acid sequence was highly homologous with those of several mannanases from B. subtilis belonging to GH family 26. Reaction temperature and pH profiles were investigated using the culture filtrate and cell-free extract of the recombinant E. coli carrying a WL-8 mannanase gene, respectively. Optimal conditions for the two fractions occurred at pH 5.5 and $60^{\circ}C$. The cell-free extract showed higher mannanase activity than the culture filtrate at above $60^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.544-549
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• 1995

The alkaline elastase is an extracellular serine protease of the alkalophilic Bacillus strain Ya-B. To increase the gene copy number and the production level of the alkaline elastase Ya-B, we designed, on the B. subtilis chromosome, a gene amplification of the 10.6 kb repeating unit containing amyE, aleE (alkaline elastase Ya-B gene) and tmrB. The aleE was inserted between amyE and tmrB, and B. subtilis APT119 strain was transformed with this amyE-aleE-tmrB-junction region fragment. As a result, we succeeded in obtaining tunicamycin-resistant (Tm$^{r}$) transformants (Tf-1, Tf-2) in which the designed gene amplification of 10.6 kb occurred in chromosome. The transformants showed high productivity of $\alpha $-amylase and alkaline elastase Ya-B. The copy number of the repeating unit (amyE-aleE-tmrB) was estimated to be 25, but plasmid vector (pUC19) was not integrated. The amplified aleE of chromosome was more stable than that of plasmid in absence of antibiotics.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.98.1-98
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• 2003

Antagonistic effect of bacterial strains isolated from phylloplane of strawberry plants grown In greenhouse was tested on Botrytis cinerea Among the promising bacterial strains, Bacillus sp. S1-0210 showed highest inhibition of mycelial growth of B. cinerea and a broad spectrum of antifungal activities against many plant pathogenic fungi in vitro. Bacillus sp. S1-0210 was identified as Bacillus subtilis based on the analysis of 185 rDNA as well as its biochemical characteristics. Application of wettable powder formulation of B. subtiiis S1-0210 significantly reduced the incidence of gray mold on trawberry fruits during storage. Results showed that treatment of B. subtilis S1-0210 decreased the incidence of gray mold by 4.8％ whereas the incidence in control was 77.9％, indicating that the formulation of B. subtilis S1-0210 will be practically applied on strawberry fruits as a biocontrol agent of gray mold during storage.

• Journal of Environmental Health Sciences
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• v.12 no.2
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• pp.67-74
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• 1986

The study was carried out to investigate for the property of Bacillus strains, on the native growthed microflora in Korean native soybean paste, and Bacillus strains of the high enzyme producing, were selected and identificated, from the microflora, that is, identificated Bacillus strains beared resemblance to B. subtills, on the colony, appearance was pellicle, surface's spreading, color creamy-thin browned, colony elevation flated, and edge lobated, the identfficated B. subtills strain named for the B. subtilis SCF. For the protease activity of B. subtilis SCF, according to the variation with pH, the pH stability was pH 7~8, and on the its protease activity, optimum temperature was 40$\circ$C, on the other hand, temperature of the highest stability of the protease was 50$\circ$C.

• Microbiology and Biotechnology Letters
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• v.51 no.1
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• pp.59-68
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• 2023

Xylanase is an industrially relevant enzyme used for the production of xylobiose and xylose. Various methods are used to enhance the microbial yield of xylanase. In the present study, co-culturing of Bacillus subtilis and Bacillus pumilus were investigated using submerged fermentation for xylanase production, which was markedly increased when sal, sagwan, newspaper, wheat bran, and xylan were used as single carbon sources. Maximum xylanase production was reported after 5 days of incubation in optimized media at pH 7.0 and 37℃, resulting in 2.69 ± 0.25 µmol/min by coculture. The 1:1 ratio of sal and sagwan in optimized production media was shown to be suitable for xylanase synthesis in submerged fermentation (SMF). In comparison to mono-culture using B. pumilus and B. subtilis, co-culturing resulted in an overall 3.8-fold and 2.15-fold increase in xylanase production, respectively.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.122-129
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• 1998

It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.