• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.552-558
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• 2021

Traditional foods are manufactured according to the characteristics of each region for the nations of the world. Korea has mainly farmed, and seasonings have developed around rice and vegetables. In fermented foods, lactic acid bacteria such as Lactobacillus sp. and Pediococcus sp. and Bacillus sp. were isolated and identified from fermented foods. In this study, lactic acid bacteria were isolated and identified from commercially available traditional Korean fermented foods, and candidate strains were selected through antibacterial activity tests on human and fish disease bacteria. Thereafter, the final strain was selected by examining the resistance to simulated gastric and intestinal fluids, and hemolysis. The three (M1, K1, C13) final selected latic acid bacteria were miced with prebiotics and the antibacterial activity of synbiotics was evaluated. As for the fist antibacterial activity result, C13 showed high antibacterial acitivity in human diseases and fish diseases. Then, M1, K1 and C13, which did not produce β-haemolysis and were resistant to simulated gastric and intestinal fluids, were subjected to the second antibacterial activity of synbiotics. When the three prebiotics (FOS, GOS, Inulin) and probiotics (M1, K1, C13) were mixed, the antibacterial activity was increased or inhibited. Based on the 16S rRNA gene sequencing results, K1 and M1 were analyzed as Bacillus tequiensis 99.72%, Bacillus subtilis 99.65%, Bacillus inaquosorum 99.72%, Bacillus cabrialesii 99.72%, Bacillus stercoris 99.58%, Bacillus spizizenii 99.58%, Bacillus halotolerans 99.58%, and Bacillus mojavensis 99.51%. And C13 was analyzed as Bacillus velezensis 99.71%, Bacillus nematocida 99.36%, Bacillus amyloliquefaciens 99.44%, Bacillus atrophaeus 99.22%, and Bacillus nakamurai 99.44%.

• Proceedings of the Korean Society of Life Science Conference
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• 2006.09a
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• pp.49.2-49.2
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• 2006

• Research in Plant Disease
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• v.29 no.4
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• pp.390-398
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• 2023

Apple white rot, caused by Botryosphaeria dothidea, is one of the important diseases in Korea. B. dothidea can cause pre- and postharvest decay on apple fruit as well as canker and dieback of apple trees. In this study, we isolated bacteria from the trunk of apple trees and tested their antagonistic activity against B. dothidea. Five bacterial isolates (23-168, 23-169, 23-170, 23-172, and 23-173) were selected that were most effective at inhibiting the mycelial growth of the pathogens. The isolate 23-172 was identified as Bacillus amyloliquefaciens and four isolates 23-168, 23-169, 23-170, and 23-173 were identified as Bacillus velezensis by RNA polymerase beta subunit (rpoB) and DNA gyraseA subunit (gyrA) gene sequencing. All isolates showed strong antagonistic activity against B. dothidiea as well as Colletotrichum fructicola and Diaporthe eres. All isolates exhibited cellulolytic, proteolytic and phosphate solubilizing activities. In particular, two isolates 23-168, 23-169 were shown to significantly reduce the size of white rot lesions in pretreated apple fruits. These results will provide the basis for the development of a fungicide alternative for the control of white rot of apple.

• Molecules and Cells
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• v.45 no.7
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• pp.502-511
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• 2022

Bacterial volatile compounds (BVCs) exert beneficial effects on plant protection both directly and indirectly. Although BVCs have been detected in vitro, their detection in situ remains challenging. The purpose of this study was to investigate the possibility of BVCs detection under in situ condition and estimate the potentials of in situ BVC to plants at below detection limit. We developed a method for detecting BVCs released by the soil bacteria Bacillus velezensis strain GB03 and Streptomyces griseus strain S4-7 in situ using solid-phase microextraction coupled with gas chromatography-mass spectrometry (SPME-GC-MS). Additionally, we evaluated the BVC detection limit in the rhizosphere and induction of systemic immune response in tomato plants grown in the greenhouse. Two signature BVCs, 2-nonanone and caryolan-1-ol, of GB03 and S4-7 respectively were successfully detected using the soil-vial system. However, these BVCs could not be detected in the rhizosphere pretreated with strains GB03 and S4-7. The detection limit of 2-nonanone in the tomato rhizosphere was 1 µM. Unexpectedly, drench application of 2-nonanone at 10 nM concentration, which is below its detection limit, protected tomato seedlings against Pseudomonas syringae pv. tomato. Our finding highlights that BVCs, including 2-nonanone, released by a soil bacterium are functional even when present at a concentration below the detection limit of SPME-GC-MS.

• Journal of the Korea institute for structural maintenance and inspection
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• v.27 no.1
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• pp.103-108
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• 2023

In this study, basic research was conducted to secure microbial resources applicable to autonomous crack healing concrete. To this end, in this experiment, biomineral-forming microorganisms were separated from natural sources, and the ability of survival in cement and calcium carbonate precipitation were compared to secure suitable microbial resources. Bacillus-type bacteria forming endospores were isolated from the sample, and the amount of calcium carbonate produced by the six microorganisms identified by 16S rRNA sequencing was compared. Two types of microorganisms, Bacillus velezensis and Bacillus subtilis, with the highest calcium carbonate precipitation were selected, and the survival of the microorganisms was confirmed through phase contrast microscopy after being cured after being added to the mortar. In addition, it was confirmed that the autonomous crack healing capability by the crack healing material produced by microorganisms was confirmed by artificially generating cracks in the mortar.

• Journal of Life Science
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• v.33 no.10
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• pp.808-819
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• 2023

This study was conducted to develop a selection marker for the identification of the Bacillus licheniformis K12 strain in microbial communities. The strain not only demonstrates good growth at moderate temperatures but also contains enzymes that catalyze the decomposition of various polymer materials, such as proteases, amylases, cellulases, lipases, and xylanases. To identify molecular markers appropriate for use in a microbial community, a search was conducted to identify variable gene regions that show considerable genetic mutations, such as recombinase, integration, and transposase sites, as well as phase-related genes. As a result, five areas were identified that have potential as selection markers. The candidate markers were two recombinase sites (BLK1 and BLK2), two integration sites (BLK3 and BLK4), and one phase-related site (BLK5). A PCR analysis performed with different Bacillus species (e.g., B. licheniformis, Bacillus velezensis, Bacillus subtilis, and Bacillus cereus) confirmed that PCR products appeared at specific locations in B. licheniformis: BLK1 in recombinase, BLK2 in recombinase family protein, and BLK3 and BLK4 as site-specific integrations. In addition, BLK1 and BLK3 were identified as good candidate markers via a PCR analysis performed on subspecies of standard B. licheniformis strains. Therefore, the findings suggest that BLK1 can be used as a selection marker for B. licheniformis species and subspecies in the microbiome.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.246-253
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• 2012

The purpose of this study was to investigate the molecular diversity of rhizobacteria associated with ginseng of varying age levels and their plant benefiting attributes. A total of 143 different isolates belonging to 15 different bacterial genera were recovered. Although variation was found in the rhizobacterial community due to age of the plant, majority of bacteria belong to Firmicutes (58%). In which, Bacillus was found to be the predominant genus irrespective of age of the ginseng. To assess the plant benefiting attributes, 30 representative isolates were selected. The results indicated that some of the isolates could exhibit multiple plant growth promoting traits like secretion of cell wall degrading enzymes, production of indole-3-acetic acid, synthesis of siderophores, solubilization of phosphates and soil pathogens inhibition. It can be suggested that strains of B. subtilis, B. amyloliquefaciens, B. velezensis, and B. licheniformis were positive for all the above traits, which have potential to be used as plant growth promoting inoculants to improve ginseng crop in the future.

• The Korean Journal of Pesticide Science
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• v.20 no.3
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• pp.189-196
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• 2016

Several microorganisms in particular Bacillus subtilis group have been isolated from diverse places such as soils and the gastrointestinal tract of ruminants etc., and used as biocontrol agent against various plant pathogens and utilized as plant growth promoting agents. Among them, Bacillus is well known as one of the most useful bacteria for biocontrol and plant growth promotion. Bacterium GH1-13 was isolated from a reclaimed paddy field in Wando Island and identified as Bacillus velezensis using phylogenetic analysis on the basis of 16S rRNA and gyrB gene. It was confirmed that GH1-13 produced indole acetic acid (IAA) associated with promoted growth of rice root. GH1-13 showed characteristics of antagonization against the main pathogen of rice as well as diverse pathogenic fungi. GH1-13 had biosynthetic genes, bacillomycin, bacilycin, fengycin, iturin, and surfactin which are considered to be associated closely with inhibition of growth of pathogenic fungi and bacteria. This study showed that GH1-13 could be used as a multifunctional agent for biocontrol and growth promotion of crop.

• Animal Bioscience
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• v.35 no.12
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• pp.1892-1903
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• 2022

Objective: A series of experiment were conducted to evaluate the effects of replacing a part of soybean meal (SBM) at 6% of broiler diets with fermented soybean meal (FSBM) obtained by single or two-stage fermentation by measuring growth performance, antioxidant activity in the jejunum and distal intestinal microflora. Methods: Soybean meal samples were prepared by single-stage fermentation using Bacillus velezensis (Bv) (FSBM_B), or Lactobacillus spp. (as commercial control) (FSBM_L). Additional SBM sample was prepared by two-stage fermentation using Bv and subsequently using Lactobacillus brevis ATCC 367 (Lb) (FSBM_B+L). Enzyme activity, chemical composition, trichloroethanoic acid-nitrogen solubility index (TCA-NSI) and antioxidant activity were measured. Then, in an in vivo study, 320 Ross308 broilers were divided into four groups with ad libitum supply of feed and water. Four groups were fed either a corn-soybean meal diet (SBM), or one of fermented SBM diets (FSBM_B+L, FSBM_B, and FSBM_L). Growth, serum characteristics, microflora, and the mRNA expression of selected genes were measured. Results: Compared to SBM, FSBM_B+L contained lower galacto-oligosaccharide, allergic protein, and trypsin inhibitor, and higher TCA-NSI by about three times (p<0.05). Reducing power and 1,1-diphenyl-2-picrylhydrazyl free radical scavenging ability correlated positively with the TCA-NSI content in FSBM. Growth performances were not significantly different among four groups. In jejunum of 35-day-old broilers, partial replacement of SBM by FSBM_B+L increased the activity of superoxide dismutase and catalase (CAT), and the FSBM_B group had the highest catalase activity (p<0.05). Partial replacement of SBM by FSBM increased relative mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and peptide transporter 1 (PepT1) (p<0.05); however, FSBM_B+L increased CAT mRNA level to 5 times of the control (p<0.05). Conclusion: Using Bv- and Lb-processed SBM through two-stage fermentation to partially replace 6% of diets will improve the gut's antioxidant activity under commercial breeding in broilers.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.343-349
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• 2019

Gul jeotgals (GJs) were prepared using solar salt aged for 3 years. One sample was fermented using starters, such as Bacillus subtilis JS2 and Tetragenococcus halophilus BS2-36 (each $10^6CFU/g$), and another sample was fermented without starters for 49 days at $10^{\circ}C$. Initial counts of bacilli and lactic acid bacteria (LAB) in non-starter GJ were found to be $3.20{\times}10^2$ and $7.67{\times}10^1CFU/g$ on day 0, and increased to $1.37{\times}10^3$ and $1.64{\times}10^6CFU/g$ on day 49. Those of starter GJ were found to be $2.10{\times}10^5$ and $3.30{\times}10^7CFU/g$ on day 49, indicating the growth of starters. The pH values of GJ were $5.93{\pm}0.01$ (non-starter) and $5.92{\pm}0.01$ (starter) on day 0 and decreased to $5.78{\pm}0.01$ (non-starter) and $5.75{\pm}0.01$ (starter) on day 49. Amino-type nitrogen (ANN) production increased continuously during fermentation, and $407.19{\pm}15.85$ (non-starter) and $398.04{\pm}13.73$ (starter) mg% on day 49. Clone libraries of 16S rRNA genes were constructed from total DNA extracted from non-starter GJ on days 7, 21, and 42. Nucleotide sequences of Escherichia coli transformants harboring recombinant pGEM-T easy plasmid containing 16S rRNA gene inserts from different bacterial species were analyzed using BLAST. Uncultured bacterium was the most dominant group and Gram - bacteria such as Acidovorax sp., Afipia sp., and Variovorax sp. were the second dominant group. Bacillus amyloliquefaciens (day 7), Bacillus velezensis (day 21 and 42), and Bacillus subtilis (day 42) were observed, but no lactic acid bacteria were detected. Acidovorax and Variovorax species might play some role in GJ fermentation. Further studies on these bacteria are necessary.