• Food Science and Preservation
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• v.24 no.8
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• pp.1168-1179
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• 2017

In this study, we tried to screen the Bacillus strain having safety probability by isolation of strains from traditional fermented food, measurement of probiotic properties, and the fermentative characteristics of Cheonggukjang. We isolated 400 Bacillus-like isolates from traditional fermented foods. Selected strains examined on the prevalent characteristic such as extracellular enzyme and antibacterial activities, and their safety probability was confirmed by biogenic amine productivity, hemolytic, and harmful substances and enzyme productivity. We selected the 5 strains by analysis of biogenic amine, antibacterial and B. cereus toxic associated gene. Five selected strains were examined on cell surface hydrophobicity, and bile and acid tolerance, and we selected the SRCM100730 as the final strain. SRCM100730 was confirmed B. amyloliquefaciens by 16S rRNA sequencing, and named the B. amyloloquefaciens SRCM100730 (KCCM11966P). Finally, we manufactured Cheonggukjang using SRCM100730 for confirmation of fermentation properties. Manufactured Cheonggukjang did not contain B. cereus, and showed that ${\gamma}$-PGA and extracellular enzyme activities were superior to commercial Chunggukjang. Amino nitrogen content was 544.02 mg% and 26 free amino acid were detected, and the bitterness-related amino acid content was lower than commercial Cheonggukjang. Especially, the amount of GABA was 3 fold higher than commercial Cheonggukjang. These results suggest that SRCM100730 have high availability in commercial probiotics market and fermented food industry.

• Food Science and Preservation
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• v.22 no.5
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• pp.727-734
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• 2015

In this study, three GRAS (generally recognized as safety) strain was isolated from Doenjang and Cheonggukjang and identified as a protease-producing microorganism, following the appearance of a clear zone around its colony when cultured on a medium containing skim milk. Based on an analysis of the nucleotide sequence of 16S ribosomal RNA, the strains wereas identified as Bacillus amyloliquefaciens and wereas therefore named Bacillus amyloliquefaciens CDD5, Bacillus amyloliquefaciens CPD4, and Bacillus amyloliquefaciens CGD3. Here, we analyzed the protease and ${\alpha}$-glucosidase inhibitory activities of the three B. amyloliquefaciens strains. Among the isolated strains, B. amyloliquefaciens CGD3 exhibited the highest protease activity (9.21 U/mL, 24 hr). The protease activities of B. amyloliquefaciens CDD5 and B. amyloliquefaciens CPD4 reached 1.14 U/mL and 8.02 U/mL, respectively, at 48 hr. The proteases from the three B. amyloliquefaciens strains showed the highest activities within a pH range of 8.0-9.0 at $50^{\circ}C$, and casein was found to be the preferred substrate on evaluating enzyme activity in the substrate specificity assay. The B. amyloliquefaciens strains exhibited maximal growth when the nutrient broth medium had an initial pH within the range of 5.0-10.0, 6-9% sodium chloride (NaCl), and 5% glucose. B. amyloliquefaciens CDD5 exhibited a low ${\alpha}$-glucosidase inhibition rate (5.32%), whereas B. amyloliquefaciens CPD4 and B. amyloliquefaciens CGD3 exhibited relatively higher inhibition rates of 96.89% and 97.55%, respectively.

• Korean Journal of Food Science and Technology
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• v.33 no.5
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• pp.567-573
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• 2001

Lactobacilli have been generally recognized as an important lactic acid bacteria present in the normal human intestinal flora. Fifty two Lactobacillus isolates were recovered from the feces of healthy Koreans whose age ranged from thirteen days after birth to 37 years. Among the isolates above, 17 isolates were tentatively identified as strains of Lactobacillus acidophilus and 3 isolates as strains of Lactobacillus casei. For their characterization, these twenty isolates were subjected to the experiments for the resistance to the artificial gastric juice, pH2.5 and bile salt. Interestingly, 3 strains survived pH 2.5 after 3 hour incubation in the artificial gastric juice with more than 75% of survival rate. L. acidophilus a-4 had the highest survival rate of 100%. Four strains including L. acidophilus a-3 grew gradually in MRS broth in the presence of the artificial bile salt. Cholesterol assimilation was also tested for the 20 isolates. The result showed that cholesterol concentration of the medium was reduced by 10 Lactobacillus isolates with more than 60%, as compared to the control. L. acidophilus a-2 had the highest reduction rate of 77%. Judging from these data obtained in vitro, some isolates ware likely to reach the lower small intestine after consumption without a significant loss of viability, suggesting that they had the potential to be developed as a probiotic culture which might lower the cholesterol level in human.

• Journal of agriculture & life science
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• v.45 no.2
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• pp.93-101
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• 2011

This study was conducted to investigate the effect of probiotic from pine needle microbes on growth performance, blood characteristics, carcass traits and economy of pigs. One hundred eighty pigs (L${\times}$Y${\times}$D, average body weight=75 kg) were grouped and housed with 15 animals per pen for 3 replications per each treatment. The ingredients and pine needle microbes mixed and fermented on anaerobic condition for 10 days to make the fermented diet. The basal diet was substituted with 0 (CON), 3 (T1), 5 (T2) and 10% (T3) of fermented diet, the pigs were fed for 42 days and slaughtered at the end of the trial to measure the carcass traits. Crude protein concentration of fermented diet on 10 day was higher (p<0.05) than that of on 0 day, while crude fiber concentration and pH were lower (p<0.05). The supplementation of fermented diet did not affect on growth performance and feed intake. Hemoglobin concentration of whole blood in T3 was higher (p<0.05) than that of CON, and hematocrit concentrations in CON and T1 were higher (p<0.05) than that of T2 and T3. The concentration of platelet was higher (p<0.05) in T1 than in T2 and T3. The CON had higher (p<0.05) total cholesterol, LDL-cholesterol and triglycerides concentrations than the treatments, whereas lower (p<0.05) HDL-cholesterol concentration. Carcass weight and dressing in T1 and T3 were higher (p<0.05) than those of CON and T2. The backfat thickness was higher (p<0.05) in CON and T2 than in T3. The carcass grade and economy improved by supplementation of fermented diet compare to CON. In conclusion, the supplementation of fermented diet using probiotics from pine needle microbes could improve health and meat quality and reduce the feed cost of pigs.

• Journal of Life Science
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• v.28 no.11
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• pp.1347-1353
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• 2018

The fermentation of non-digestible soy meal can convert polysaccharides into many compounds that have a wide variety of biological functions. Bacillus strains are capable of hydrolyzing non-digestible saccharides, such as melibiose, raffinose, and stachyose, found in soy meal components. A highly active ${\alpha}$-galactosidase (${\alpha}$-d-galactoside galactohydrolase, EC 3.2.1.22) was isolated from a bacterium in a traditional Korean fermented medicinal herb preparation. The isolate, T2-16, was identified as Bacillus coagulans based on its 16S rRNA sequence and biochemical properties, and the strain was named Bacillus coagulans NRR-1207. When incubated in 10%(w/v) skim milk, Bacillus coagulans NRR1207 caused a decrease in the pH of the culture medium, as well as an increase in titratable acidity and viable cell counts. This strain also showed higher activities of ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase, naphthol-AS-BO-phosphohydrolase, and acid phosphatase when compared to other enzymes. It hydrolyzed oligomeric substrates, such as raffinose and stachyose, and liberated galactose, indicating that the Bacillus coagulans NRR1207 ${\alpha}$-galactosidase hydrolyzed the ${\alpha}$-1,6 glycoside linkage. These results suggest that the decreased stachyose and raffinose contents observed in fermented soy meal are due to this ${\alpha}$-galactosidase activity. Bacillus coagulans NRR1207 therefore has potential probiotic activity and could be utilized in feed manufacturing, as well as for hydrolyzing non-digestible soy meal components.

• 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%.

• Korean Journal of Poultry Science
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• v.50 no.3
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• pp.171-185
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• 2023

This study was conducted to investigate the effects of supplementation of Allium hookeri (AH) root powder on the gut microbiome, immunity, and health in broiler chickens fed experimental diets from d 10 to 28. A total of 60 10-day-old Ross 308 broilers were weighed and assigned to two dietary treatments with 5 birds per cage in a randomized complete block design based on body weight. The two experimental diets consisted of a control diet based on corn-soybean meal and the control diet supplemented with 0.3% AH root powder. All birds were fed ad libitum with experimental diets and water for 18 d. At 28 d, two birds near the median weight from each cage were selected for cecal content and small intestinal tissue sample collection. The addition of AH changed the gut microbiome by increasing probiotic candidate beneficial bacteria such as Enterococcaceae, Lactobacillaceae, Limosilactobacillus, Cuneatibacter, and Ruminoccoides. Regarding gut immunity, the supplementation of AH resulted in changes in intestinal immune cells, including reduced CD3+CD4+ T cells, which are a type of helper T cell, in the small intestine of birds (P=0.049). Additionally, there was a tendency to increase the expression of antioxidant function-related gene such as GPX2 (P=0.060), but no significant changes were observed in cytokines such as IL1b, IL6, and IL10. Overall, the addition of AH root powder may have positive effects on the microbiome of the chickens. This may help promote gut health in broiler chickens at the age of d 10 to 28.

• Korean Journal of Poultry Science
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• v.37 no.1
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• pp.51-62
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• 2010

This study was conducted to investigate the effects of dietary supplementation of multiple probiotics on the performance, small intestinal microflora and immune response in laying hens and broilers. In Exp.1, a total of 800, 82 wk old Hy-line Brown$^{(R)}$ laying hens were assigned to one of the following five dietary treatment; Control, Antibiotics (avilamycin 6 ppm), Probiotics; PB-M (Micro-ferm$^{(R)}$ 0.2%), PB-L (Lacto-sacc$^{(R)}$ 0.1%), PB-Y (Y University probiotics 0.2%). Each treatment was replicated eight times with 20 birds in each replicate and two birds were housed in each cage. Twenty birds units were arranged according to completely randomized block design. Feeding trial lasted 6 wk under 16 h lighting regimen. The Exp. 2, was conducted with a total of 1,000 broilers chicks (Ross$^{(R)}$). They were divided into five treatments, same as those of Exp. 1. Birds were fed starter (0~3 wk) and grower (4~5 wk) diets. Each treatment was replicated four times with 50 birds per pen comprising of deep litter. In Exp. 1, egg production parameters, such as hen-day and hen-house egg production, egg weight, broken and soft shell egg production, feed intake and feed conversion were not significantly different among treatments. However, strength and thickness of eggshell were significantly (P<0.05) different. Among the probiotics, PB-Y showed the highest strength and thickness of eggshell. Eggshell color, egg yolk color and Haugh unit were not significantly influenced. In Exp. 2, overall weight gain (0~5 wk) and mortality were not significantly different among treatments. However, weight gain of birds from PB-Y treatment during starter (0~3 wk) was significantly lower than the birds from Control and Antibiotic treatment. During the whole period (0~5 wk), birds from Antibiotics treatment had higher feed intake and Production Index (PI) and lower feed conversion than birds from Control treatment. Probiotics treatments were not significantly different from the Control on feed intake and feed conversion. In Exp.1, there were significant (P<0.05) differences in leukocytes parameters, such as white blood cell (WBC), hetrophil (HE), lymphocytes (LY), monocyte (MO), eosinophil (EO) and stress index (SI; HE/LY) in the blood of layers. Birds from Antibiotics and probiotics treatments tended to increase these parameters. In Exp. 2, however, only SI was significantly (P<0.05) decreased in Antibiotics treatments. Concentration of serum immunoglobulin (IgG) were higher (P<0.05) in PB-M and PB-Y treatments when compared with Control treatment in Exp. 1. The population of E. coli significantly (P<0.05) decreased in birds from Antibiotics, PB-L and PB-Y treatments when compared with birds from Control treatment in Exp. 1. Metalbolizability of crude fat decreased significantly (P<0.05) in birds from probiotic treatments in Exp. 2. It was concluded that the response of probiotics on the productivity of layers and broilers were different. Probiotics increased strength and thickness of eggshell in layers, and decreased feed conversion and increased PI in broilers. Leukocytes and IgG tended to increase by supplementation of antibiotics and probiotics in layers. Intestinal E. coli tended to decrease in layers. Digestibility of crude fat of diet decreased in probiotics treatments broilers. Parameters of blood and microbial were more sensitive in layers than broilers.