• Asian-Australasian Journal of Animal Sciences
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• v.24 no.8
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• pp.1120-1127
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• 2011

A feeding trial was conducted to investigate the effect of dietary supplementation of Bacillus subtilis LS 1-2 grown on citrus-juice waste and corn-soybean substrate on growth performance, nutrient retention, caecal microbial population and intestinal morphology in broilers. Three hundred twenty d-old Ross chicks were randomly allotted to 4 treatments on the basis of BW in a randomized complete block design. Each treatment had 4 replicates of 20 chicks in each. Experimental diets were fed in 2 phases, starter (d 0 to 21) and finisher (d 21 to 35). Dietary treatments were; negative control (NC: basal diet without any antimicrobial), positive control (PC: basal diet added with 20 mg/kg Avilamycin), basal diet added with 0.30% Bacillus subtilis LS 1-2 grown on corn-soybean substrate (P1), and basal diet added with 0.30% Bacillus subtilis LS 1-2 grown on citrus-juice waste substrate (P2). Overall BW gain, feed intake and FCR were better (p<0.05) in PC, P1 and P2 treatments as compared to NC. Moreover, overall BW gain and FCR in PC and P2 treatments were greater than P1. Retention of CP, GE (d 21, d 35) and DM (d 35) were increased (p<0.05) in treatments PC, P1 and P2 compared with NC. At d 35, caecal Clostridium and Coliform counts were lower (p<0.05) in treatments PC, P1 and P2 than NC. Moreover, Clostridium and Coliform counts in treatment PC was lower (p<0.05) than P1. Villus height and villus height to crypt depth ratio in both duodenum and ileum were increased (p<0.05) in treatments PC, P1, P2 as compared to NC. However, retention of nutrients, caecal microbial population and intestinal morphology remained comparable among treatments P1 and P2. It is concluded that Bacillus subtilis LS 1-2 inclusion at 0.30% level had beneficial effects on broilers' growth performance, nutrient retention, caecal microflora and intestinal morphology. Additionally, citrus-juice waste can be used as substrate for growth of probiotic Bacillus subtilis LS 1-2.

• Korean Journal of Food Science and Technology
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• v.43 no.6
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• pp.735-741
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• 2011

Bealmijang is a short-term fermented regional product that is prepared with soybean and extra ingredients. In this study, starter strain candidates were screened from Bealmijang for fermented soybean paste products. Twenty one bacterial strains producing extracellular enzymes (amylase, cellulase, protease, xylanase and lipase) were isolated from Bealmijang, buckwheat sokseongjang. The isolates were assessed for fibrinolytic and antibacterial activities, and salt tolerance. Strain HJ18-4, identified as Bacillus subtilis (AB601598) by biochemical properties (89.6%) and 16S rDNA sequencing (100%), showed the highest enzymatic, fibrinolytic, and antibacterial activities among the isolates. Although the growth of HJ18-4 was inhibited by the increase of NaCl concentration, the growth still exceeded that of B. subtilis KACC 10114 at 5% and 10% NaCl. These results suggest that B. subtilis HJ18-4 is suitable as a starter for soybean paste manufacture.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.5
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• pp.736-745
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• 2016

Cheonggukjang, a traditional Korean fermented soy food, was prepared by inoculation of Bacillus subtilis EMD4 or Bacillus amyloliquefaciens EMD17 with anti-bacterial or anti-fungal activities into soybeans. Cheonggukjang was also prepared by co-inoculation of EMD4 and EMD17 (1:1, v/v). Control cheonggukjang was prepared by using B. subtilis KACC16450 (Natto strain). Growth of B. cereus cells spiked with starter organisms was completely inhibited by B. amyloliquefaciens EMD17 after 12 h of fermentation at $37^{\circ}C$. Growth of B. cereus was also inhibited by B. subtilis EMD4, but the degree of inhibition was weaker. After 48 h of fermentation, cheonggukjang samples were stored for 10 days at $4^{\circ}C$. B. cereus cells were not detected from cheonggukjang inoculated with EMD4, whereas significant numbers still present in control. The pH values of cheonggukjang samples were not significantly different. During fermentation, cheonggukjang fermented with EMD17 showed the highest fibrinolytic activity and during storage, cheonggukjang fermented with a Natto strain was the highest. Cheonggukjang fermented with a Natto strain also showed the highest amount of total phenolic compounds. The results show that control of B. cereus in cheonggukjang is possible by using starters such as B. amyloliquefaciens EMD17.

• Journal of Microbiology and Biotechnology
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• v.34 no.7
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• pp.1443-1451
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• 2024

Weizmannia coagulans can be used as a starter strain in fermented foods or as a probiotic. However, it is salt-sensitive. Here, W. coagulans genomes were compared with the genomes of strains of Bacillus species (B. licheniformis, B. siamensis, B. subtilis, and B. velezensis) that were isolated from fermented foods and show salt tolerance, to identify the basis for the salt-sensitivity of W. coagulans. Osmoprotectant uptake (Opu) systems transport compatible solutes into cells to help them tolerate osmotic stress. B. siamensis, B. subtilis, and B. velezensis each possess five Opu systems (OpuA, OpuB, OpuC, OpuD, and OpuE); B. licheniformis has all except OpuB. However, W. coagulans only has the OpuC system. Based on these findings, the opuA and opuB operons, and the opuD and opuE genes, were amplified from B. velezensis. Expression of each of these systems, respectively, in W. coagulans increased salt-tolerance. W. coagulans expressing B. velezensis opuA, opuD, or opuE grew in 10.5% NaCl (w/v), whereas wild-type W. coagulans could not grow in 3.5% NaCl. The salt resistance of B. subtilis was also increased by overexpression of B. velezensis opuA, opuB, opuD, or opuE. These results indicate that the salt-susceptibility of W. coagulans arises because it is deficient in Opu systems.

• Journal of Food Hygiene and Safety
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• v.14 no.4
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• pp.346-351
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• 1999

To develop a lactic starter to produce antimicrobial substance for inhibiting the growth of a variety of foodborne spoilage bacteria in fermented foods, we investigated the anti-bacterial effect of the antibacterial substance, produced by Lactobacillus amylovorus IMC-1, against foodborne spoilage strains, and its sensitivity on the treatment of proteolytic enzymes. L. amylovorus IMC-1, which was isolated from a traditional cheese in Inner Mongolia, produced a maximum amount of antibacterial substance in the skim milk medium after 72 h incubation at 37$^{\circ}C$, and further incubation resulted in the same activity. The substance obtained from gel filtration inhibited all strains used such as Bacillus subtilis IFO 3025, Staphylococcus aureus IAM 1011, Listeria monocytogenes VTU 206, Escherichia coli RB, and Pseudomonas fragi IFO 3458 at the concentration of 20 units/ml. This substance was found to show bactericidal action against B. subtilis, E. coli, and Ps. fragi, and bacteriostatic activity against both Staph. aureus and L. monocytogenes. The bactericidal action was due to cellular Iysis. The substance is not organic acid, hydrogen peroxide and proteinaceous compound.

• The Korean Journal of Food And Nutrition
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• v.26 no.4
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• pp.753-758
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• 2013

This study evaluated the quality characteristics for optimization of premix-type Buckwheat (Fagopyrum esculentum) Soksungjang (BS). First, according to the amount of salt (8, 10, 12%), coliform counts were not detected in BS with 10 or, 12% salt at 21 days and Bacillus subtilis HJ 18-4 as a starter at 7 days. Therefore, 10% salt with starter culture might be considered as suitable conditions for safety. Second, according to the ratio of water (1:1, 1:1.5, 1:1.8), BS prepared with the same weights of meju powder and water showed the highest aminotype nitrogen content (688.76mg%) at 35 days; thus, an adequate fermentation state was established at this condition compared to others. Third, according to the type of container (pot, plastic, glass), the aminotype nitrogen contents were higher in the plastic container and pot than in the glass bottle. Lastly, according to the batch size (1.25, 2.5 kg), the coliform of the 2.5 kg sample decreased faster than that of 1.25 kg samples at 14 days. These results suggest that BS prepared with the same ratio of water and meju powder fermented with HJ 18-4 as a starter and 10% salt in a pot, plastic container (3 weeks) and glass container (4 weeks) showed desirable fermentation qualities.

• Journal of Life Science
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• v.23 no.4
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• pp.560-568
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• 2013

Chungkookjang was fermented by B. subtilis MC31, a ${\gamma}$-amino butyric acid (GABA) producing microorganism. The characteristics of Chungkookjang were investigated while fermenting. Twenty four amino acids were detected in Chungkookjang, leucine was the highest of them all. Total cell populations of B. subtilis MC31 phase were between log $9.52{\pm}0.5$ ~ log $9.049{\pm}0.5$ CFU/g at stationary phase. Contents of moisture, crude ash, crude protein, crude lipid and crude fiber are $61.07{\pm}0.01%$, $1.52{\pm}0.01%$, $17.66{\pm}0.04%$, $8.96{\pm}0.03%$ and 2.61%, respectively. Contents of ammonia type nitrogen, amino type nitrogen and reducing sugar were increased during fermentation at $40^{\circ}C$ for 72 hr, however those of titratable acidity and total sugar were decreased. pH was slowly alkalized during fermentation. Viscous substance and protease contents in Chungkookjang were $4.7{\pm}0.05%$ and $0.519{\pm}7.36$ g/l, apiece. When the fibrin plate and Robbin method for fibrinolytic activity were applied, B. subtilis MC31 showed high activity. These results suggested that B. subtilis MC31 is suitable to be used as a starter to enhance the quality of Chungkookjang.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.419-426
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• 2015

The aims of this study were to isolate spore-forming Bacillus strains that exhibit high digestibility and anti-pathogenic bacteria toward feed for calves. Total 136 spore-forming strains were isolated from finished feeds and their ingredients. Among them, 93 strains were identified as Bacillus species when analyzed by 16S rRNA sequencing. For industrial use, three strains named as Bacillus licheniformis SHS14, B. subtilis LCB7, B. amyloliquefaciens LCB10 were selected after evaluating the industrial standards that are related with heat and acid resistance, enzyme activities, and anti-pathogenic activities against Samonella dublin ATCC15480 and E. coli K99. After each culture, 3 selected strains were mixed together at 1:1:1 (v/v/v) ratio and then prepared as the mixed starter culture for feeding. The changes in microbial community were analyzed via 16S rRNA metagenomics. The initial community ratio among three strains was maintained even after manufacturing into final products. Also, in vitro, enzymatic and anti-pathogenic activities were almost same as those when cultured in single culture, and results of anti-pathogenic activities conducted with calves showed 90% activities against lincomycin, which would be indicative of a promising feed starter.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2090-2099
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• 2015

Recently, the cardiovascular disease has been widely problematic in humans probably due to fibrin formation via the unbalanced Western style diet. Although direct (human plasmin) and indirect methods (plasminogen activators) have been available, bacterial enzyme methods have been studied because of their cheap and mass production. To detect a novel bacterial fibrinolytic enzyme, 111 bacterial strains with fibrinolytic activity were selected from kimchi. Among them, 14 strains were selected because of their stronger activity than 0.02 U of plasmin. Their 16S rRNA sequence analysis revealed that they belong to Bacillus, Leuconostoc, Propionibacterium, Weissella, Staphylococcus, and Bifidobacterium. The strain B. subtilis ZA400, with the highest fibrinolytic activity, was selected and the gene encoding fibrinolytic enzyme (bsfA) was cloned and expressed in the E. coli overexpression system. The purified enzyme was analyzed with SDS-PAGE, western blot, and MALDI-TOF analyses, showing to be 28.4 kDa. Subsequently, the BsfA was characterized to be stable under various stress conditions such as temperature (4-40oC), metal ions (Mn²⁺, Ca²⁺, K²⁺, and Mg²⁺), and inhibitors (EDTA and SDS), suggesting that BsfA could be a good candidate for development of a novel fibrinolytic enzyme for thrombosis treatment and may even be useful as a new bacterial starter for manufacturing functional fermented foods.

• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.400-405
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• 2008

The effects of different starter cultures on the fermentative characteristics of cheonggukjang were examined by using three Bacillus strains. The strains included Bacillus subtilis KCTC 1021 as a control, Bacillus sp. Kn-10 (Kn-10) isolated from a commercial cheonggukjang, and Bacillus sp. B-59 (B-59) isolated from rice straw. There were no significant differences in pH or viable cells among the different cheonggukjang samples during fennentation for 72 hr at $40^{\circ}C$. However, the sample prepared with B-59 had higher slime content and protease activity than the controls and Kn-10 samples. DPPH free radical scavenging activity was higher in the B-59 sample and lower in the control and Kn-10 samples when compared to steamed soybeans after fermentation for 72 hr at $40^{\circ}C$. The total amino acid contents the cheonggukjangs were 34869.98 mg% (B-59), 34481.89 mg% (control), and 31791.09 mg% (Kn-10). Glutamic acid and lysine contents were higher in the B-59 sample than in the control. Finally, the cheonggukjang fermented using the B-59 strain had improved sensory qualities such as color, taste, texture, and overall acceptability compared to the control and Kn-10 samples.