• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.5
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• pp.600-605
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• 2006

B. subtilis JM3 pretense from naturally fermented anchovy sauce was purified in $40{\sim}60%$ ammonium sulfate fraction. In order to accelerate the fermentation of anchovy sauce,2% and 4% of crude B. subtilis JM3 protease were added to 6 month-fermented anchovy sauces, respectively and then the various quality characteristics such as pH, lactic acid, amino-nitrogen, VBN, browning and hydrolysis degree, VBN, and sensory evaluation were analyzed at different storage times. pH was constant during storage time in all samples, whereas lactic acid contents of anchovy sauces hydrolyzed by 2% and 4% proteases were higher than that of control. The amino-nitrogen and volatile basic nitrogen contents of anchovy sauce with 2% and 4% proteases were twice higher than those of control. Anchovy sauces with 2% and 4% pretense increased the hydrolysis rate by 27% and 32%, respectively. Browning degree of anchovy sauce with 4% was higher than those of 2% and control. Anchovy sauce with 2% and 4% proteases was good in sensory evaluation of color, aroma, and taste attributes.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.398-409
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• 2018

In the present study, biogenic amine-forming Bacillus spp. and bacteriocin-producing lactic acid bacteria (LAB) isolated from Doenjang were generally identified through 16S rRNA gene sequencing, and the physicochemical and microbiological characteristics of cheonggukjang prepared using the isolated strains were investigated. Biogenic amine-producing bacteria from the samples were identified as Bacillus licheniformis DB102, B. subtilis DB203, B. stearothermophilus DB206, B. pumilus DB209, B. subtilis DB310, B. coagulans DB311, B. cereus DB313, B. amyloliquefaciens DB714, B. amylolique-faciens DB915, B. licheniformis DB917, B. cereus DB1019, B. subtilis DB1020, B. megaterium DB1022. The bacteriocin-producing LAB showed antibacterial effect against biogenic amine-producing Bacillus spp. were identified as Lactobacillus plantarum DLA205, L. brevis DLA501, L. fermentum DLA509, L. acidophilus DLA703, and Enterococcus faecalis DLA804. The bacteriocin produced by the LAB significantly decreased the viable numbers and the amine production ability of the biogenic amine-forming Bacillus spp. in a concentration dependent manner. Therefore, the pH, ammonia nitrogen and biogenic amine content of cheonggukjang prepared by mixed culture of the LAB and Bacillus spp. were significantly decreased compared to the control group.

• The Plant Pathology Journal
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• v.32 no.2
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• pp.145-156
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• 2016

Fusarium graminearum Schwabe causes Fusarium head blight (FHB), a devastating disease that leads to extensive yield and quality loss of wheat and other cereal crops. Twelve isolates of F. graminearum were collected from naturally infected spikes of wheat from Assiut Egypt. These isolates were compared using SRAP. The results indicated distinct genetic groups exist within F. graminearum, and demonstrated that these groups have different biological properties, especially with respect to their pathogenicity on wheat. There were biologically significant differences between the groups; with group (B) isolates being more aggressive towards wheat than groups (A) and (C). Furthermore, Trichoderma harzianum (Rifai) and Bacillus subtilis (Ehrenberg) which isolated from wheat kernels were screened for antagonistic activity against F. graminearum. They significantly reduced the growth of F. graminearum colonies in culture. In order to gain insight into biological control effect in situ, highly antagonistic isolates of T. harzianum and B. subtilis were selected, based on their in vitro effectiveness, for greenhouse test. It was revealed that T. harzianum and B. subtilis significantly reduced FHB severity. The obtained results indicated that T. harzianum and B. subtilis are very effective biocontrol agents that offer potential benefit in FHB and should be harnessed for further biocontrol applications. The accurate analysis of genetic variation and studies of population structures have significant implications for understanding the genetic traits and disease control programs in wheat. This is the first known report of the distribution and genetic variation of F. graminearum on wheat spikes in Assiut Egypt.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.131-137
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• 2001

Previous work has identified that only the catabolite responsive element A (creA; previously called cre-2) out of two potential cre sequences (cre-1: nucleotide +160 to +173 and cre-2: +173 to +186), recognized within the coding region of the xylanaseA gene (xynA) of Bacillus stearothermophilus No.236, was actually, was actually involved in the carbon catabolite repression(CCR) of xynA expression in B. subtilis. However, the level of CCR of xynA expression in the original B.stearothermophilus No.236 strain (70-fold repression). Therefore, to search for an additional cre element in the promoter region, the upstream region of the xynA gene was subcloned by chromosome walking, and as a result, another potential cre element (nucleotide -124∼-137; designated creB) was recognized in this region. The cre-like sequence revealed a high homology to the cre consensus sequence. The xylanase activity of B. subtilis MW15 bearing pWPBR14 (containing creA and creB) cultured in a medium containing xylose as the sole carbon source was about 7.7 times higher than that observed for the same culture containing glucose. B. subtilis MW15 bearing pWPBR23 (containing only creA) produced an activity about 2.4 times higher. This pattern of CCR was confirmed using derivatives of xynA::aprA fusion plasmids. Furthermore, a measurement of the amounts of the xynA transcript showed a similar pattern as that for the production of xylanase. In addition, the synthesis of xylanase in B. subtilis QB7115 [a catabolite control protein A (ccpA) mutant strain] carrying pWPBR14 was almost completely relieved from glucose repression. Together, these results lead to a conclusion that the CCR of the expression of the xynA gene is mediated by CcpA binding at creA and creB sites in B. subtilis.

• Applied Biological Chemistry
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• v.7
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• pp.53-58
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• 1966

A study was carried out to investigate the microorganisms and their floral changes during the tobacco fermentation. The results were sumarized as follows. I. The molds in the "tobacco leaves" were isolated and identified as follows; Aspergllus flavus Link, A. restrictus G. Smith, A. nidulans (Eidam) Wint, A. awamori, Oidium sp. Edmundmasonia sp. Spicaria sp. II. The bacteria in the tobacco were isolated and identified as follows; Bacillus subtilis, B. subtilis var aterriums. B. licheniformis, B. cereus, B. Pumilus, B. megaterium, Flavobacterium harrisonii, Aerobacter aerogenes. III. The counts of the microorganisms on leaves taken from bulks of the fermenting leaf tobacco revealed the presence of relatively small number on the initial stage of the fermentation. During the tobacco fermentation the number of molds increase gradually to the maximum until the 14 th. day of the fermenation, followed by showing, the plateau, and the bacteria population revealed the maximum on the 7 th. day, then declined slowly.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1863-1870
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• 2015

Fibrinolytic enzyme genes (aprE2, aprE176, and aprE179) were introduced into the Bacillus subtilis 168 chromosome without any antibiotic resistance gene. An integration vector, pDG1662, was used to deliver the genes into the amyE site of B. subtilis 168. Integrants, SJ3-5nc, SJ176nc, and SJ179nc, were obtained after two successive homologous recombinations. The integration of each fibrinolytic gene into the middle of the amyE site was confirmed by phenotypes (Amy-, Spec^S) and colony PCR results for these strains. The fibrinolytic activities of the integrants were higher than that of B. subtilis 168 by at least 3.2-fold when grown in LB broth. Cheonggukjang was prepared by inoculating each of B. subtilis 168, SJ3-5nc, SJ176nc, and SJ179nc, and the fibrinolytic activity of cheonggukjang was 4.6 ± 0.7, 10.8 ± 0.9, 7.0 ± 0.6, and 8.0 ± 0.2 (U/g of cheonggukjang), respectively at 72 h. These results showed that construction of B. subtilis strains with enhanced fibrinolytic activities is possible by integration of a strong fibrinolytic gene via a marker-free manner.

• The Korean Journal of Mycology
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• v.42 no.4
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• pp.333-340
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• 2014

The aim of this study was to isolate a potential multifunctional biocontrol agent from bacteria for control of multiple plant diseases as an alternative to fungicides. A total of 201 strains were isolated from soil undamaged by repeated cultivation in Sunchang and their ability to produce antibiotics, siderophores and extracellular enzymes such as protease, cellulase and amylase was investigated. Selected strain SCS3 produced cellulose, protease and amylase. This strain also produced siderophores and showed excellent antifungal activity against various phytopathogens. SCS3 was identified as Bacillus subtilis using 16S rRNA sequencing, and named Bacillus subtilis SCS3. Finally, physiological and biochemical characteristics of B. subtilis SCS3 were examined. From the results, B. subtilis SCS3 was found to be a useful multifunctional biocontrol agent against various phytopathogens.

• Journal of Life Science
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• v.11 no.1
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• pp.42-46
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• 2001

Subtilisin YaB, produced by alkalophilic Bacillus strain YaB, is an extracellular alkaline serine protease having 55% homology to subtilisin BPN'. It is synthesized as a 378-amino acid preproenzyme and secreted into the culture medium as a 265-amino acid mature protease. To examine the role of pro-sequence for the secretion of subtilisin YaB, we have studied the expression, in Bacillus subtilis, of a mutant preprosubtilisin YaB in which active site Ser214 is substituted with Cys. The use of a six protease-deficient strain, WB600, was required for its efficient production. The prosubtilisin YaB, thus produced, was indeed secreted into the culture medium and was processed to its mature form upon treatment with exogenously added active subtilisin YaB. From these results, we have concluded that the processing of pro-sequence is not essential for the secretion of the enzyme.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.311-317
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• 2006

Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

• The Korean Journal of Pesticide Science
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• v.14 no.1
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• pp.49-53
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• 2010

Disease control efficacy was evaluated with use of Bacillus subtilis KB-401 against cucumber powdery mildew in a greenhouse and fields. B. subtilis KB-401 showing inhibitory effect on mycelial growth of various phytopathogenic fungi was formulated for the evaluation. The formulated biofungicide of B. subtilis KB-401 was less effective at 1,000 times dilution rate than that at 250 or 500 times dilution rate. The powdery mildew was successfully controlled by the biofungicide at the early stage of disease development. The field performance of the biofungicde was conducted in Asan and Cheonan city. Three or four consecutive applications of the biofungicide at 500 dilution rate with 10-day intervals resulted in considerable efficacy of disease control as high as 83.3%.