• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.5
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• pp.266-273
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• 2020

This study was undertaken to determine the effects of diet supplementation with complex probiotics (CPB), on growth performance, nutrient digestibility, blood metabolites, noxious gas, and fecal microflora in weaning pigs. On the basis of body weight, a total of 234 weaned pigs (Landrace×Yorkshire×Duroc, 6.14±0.78kg) were randomly allotted to 3 treatments and 6 replicates (13 pigs per pen). The experimental diets were fed in a meal form for 28 days (days 0-14, PhaseI, and days 15-28, PhaseII). The dietary treatment groups were as follows: T1 (basal diet), T2 (T1+0.13% CPB) and T3 (T1+0.25% CPB). The CPB supplement contained Bacillus subtilis 1.0×10⁶ CFU/g, Enterococcus faecium 1.0×10⁶ CFU/g, Saccharomyces cerevisiae 1.0×10⁶ CFU/g, Bacillus licheniformis 3.0×10⁸ CFU/g, and Bacillus polyfermenticus 3.0×10⁸ CFU/g. Pigs fed the T3 diet showed an increase (p<0.05) in the overall average daily gain and average daily feed intake, increased (p<0.05) crude protein digestibility in PhaseI, and greater (p<0.05) dry matter and gross energy digestibility in PhaseII. Supplementation of CPB had no effect on the blood profile. Furthermore, pigs fed the T3 diet had lower (p<0.05) NH₃ emission and overall count of fecal Clostridium spp. In conclusion, we believe that CPB supplementation has a beneficial effect on growth performance, nutrient digestibility, noxious gas, and fecal microflora in weaning pigs.

• Proceedings of the Microbiological Society of Korea Conference
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• 1991.04a
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• pp.225-236
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• 1991

The genes encoding the thermostable $\alpha$-amylase and maltogenic amylase from Bacillus lichenciformis were cloned and expressed in E. coli. The recombinant plasmid pTA322 was found to contain a 3.1kb EcoRI genomic DNA fragment of the thermostable $\alpha$-amylase. The cloned $\alpha$-amylase was compared with the B. licheniformis native $\alpha$-amylase. Both $\alpha$-amylase have the same optimal temperature of $70^{\circ}C$ and are stable in the pH range of 6 and 9. The complete nucleotide sequences of the thermostable $\alpha$-amylase gene were determined. It was composed of one open reading rame of 1,536 bp. Start and stop codons are ATG and TAG. From the amino acid sequence deduced from the nucleotide sequence, the cloned thermostable $\alpha$-amylase is composed of 483 amino acid residues and its molecular weight is 55,200 daltons. The content of guanine and cytosine is $47.46mol\%$ and that of third base codon was $53_41mol\%$. The recombinant plasmid, pIJ322 encoding the maltogenic amylase contains a 3.5kb EcoRI-BamHI genomic DNA fragment. The optimal reaction temperature and pH of the maltogenci amylase were $50^{\circ}C$ and 7, respectively. The maltogenic amylase was capable of hydrolysing pullulan, starch and cyclodextrin to produce maltose from starch and panose from pullulan. The maltogenic amylase also showed the transferring activity. The maltogenic amylase gene is composed of one open reading frame of 1,734bp. Start and stop codons are ATG and ATG. At 2bp upstream from start codon, the nucleotide sequence AAAGGGGGAA seems to be the ribosome-binding site(RBS, Shine-Dalgarno sequence). A putative promoter(-35 and-10 regions) was found to be GTTAACA and TGATAAT. From deduced amino acid sequence from the nucleotide srquence, this enzyme was comosed of 578 amino acid residues and its molecular weight was 77,233 daltons. The content of guanine and cytosine was $48.1mol\%$. The new recombinant plasmid, pTMA322 constructed by inserting the thermostable $\alpha$-amylase gene in the EcoRI site of pIJ322 to produce both the thermostable $\alpha$-amylase and the maltogenic amylase were expressed in the E. coli. The two enzymes expressed from E. coli containing pTMA322 was reacted with the $15\%$ starch slurry at $40^{\circ}C$ for 24hours. The distribution of the branched oligosaccharides produced by the single-step process was of the ratio 50 : 50 between small oligosaccharide up DP3 and large oligosaccharide above DP3.

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1717-1725
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• 2013

Traditional soybean paste from Shandong Liangshan and Tianyuan Jiangyuan commercial soybean paste were chosen for analysis and comparison of their bacterial and fungal dynamics using denaturing gel gradient electrophoresis and 16S rRNA gene clone libraries. The bacterial diversity results showed that more than 20 types of bacteria were present in traditional Shandong soybean paste during its fermentation process, whereas only six types of bacteria were present in the commercial soybean paste. The predominant bacteria in the Shandong soybean paste were most closely related to Leuconostoc spp., an uncultured bacterium, Lactococcus lactis, Bacillus licheniformis, Bacillus spp., and Citrobacter freundii. The predominant bacteria in the Tianyuan Jiangyuan soybean paste were most closely related to an uncultured bacterium, Bacillus licheniformis, and an uncultured Leuconostoc spp. The fungal diversity results showed that 10 types of fungi were present in the Shandong soybean paste during the fermentation process, with the predominant fungi being most closely related to Geotrichum spp., an uncultured fungal clone, Aspergillus oryzae, and yeast species. The predominant fungus in the commercial soybean paste was Aspergillus oryzae.

• Horticultural Science & Technology
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• v.32 no.6
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• pp.879-886
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• 2014

This study was conducted to test in vitro the antagonistic effect of composted liquid manure (CLM) against soil-borne turfgrass pathogenic fungi, Rhizoctonia solani AG-2-2 (IIIB) (brown patch), R. solani AG-2-2 (IV) (large patch), and Sclerotinia homoeocarpa (dollar spot) for environmentally friendly turfgrass management. CLMs were collected from 9 livestock excretion treatment facilities around the country including Gunwi (GW), Hapcheon (HC), Hoengseong (HS), Icheon (IC), Iksan (IS), Muan (MA), Nonsan (NS), and Yeoju (YJ). CLMs of IC, GW, and IS showed s ignificant (p < 0.05) mycelium growth inhibition that was 17.8%, 20.4%, and 48.0% against R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. A t otal of 110 bacterial isolates were obtained from the CLMs that showed antagonistic effects. Among them, 5, 4, and 10 microbe isolates showed promising antifungal activity against mycelium growth of R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. The bacterial isolates ICIIIB60, GWIV70, and ISSH20 effectively inhibited the mycelial growth of three soil-borne turfgrass pathogens. Selected bacterial isolates were identified as Alcaligenes sp., Bacillus licheniformis Ab2, and B. subtilis C7-3 through 16s rDNA gene sequence analysis. Among 5 fungicides, the most compatible fungicide with ICIIIB60, GWIV70, and ISSH20 was tebuconazol, toclofos-methyl and toclofos-methyl, respectively. These findings suggested that CLMs could be effectively used not only as organic liquid fertilizer sources but also as biological control agents for soil-borne turfgrass diseases such as brown patch, large patch, and dollar spot.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.2
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• pp.171-180
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• 2017

Objective: This study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR) silage. Methods: The TMRs were prepared with soybean curd residue, alfalfa hay (ATMR) or Leymus chinensis hay (LTMR), corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties. Results: Both ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens), B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens) in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively. Conclusion: The microbial amylase contributes to starch hydrolysis during the ensiling process in both TMR silages, whereas the microbial hemicellulase participates in the hemicellulose degradation only at the early stage of ensiling.

• Food Science and Preservation
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• v.21 no.6
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• pp.866-877
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• 2014

The aim of this study was to develop a new functional traditional fermented soybean food using Cudrania tricuspidata fruits and fermentation microbes isolated from traditional fermented food. Aspergillus oryzae koji, Lactobacillus sp., and Bacillus sp. were used for the selection of a suitable microbe for the fermentation of Cudrania tricuspidata fruits, and as a result, Bacillus licheniformis SCDB 1234 was selected. SCDB 1234 enhanced the concentration of kaempferol in the Cudrania tricuspidata fruits from 9.54 to $217.04{\mu}g/g$ (about 22 times). The DPPH radical scavenging activity of the fermented materials was similar to that of BHA and BHT (92~99 ppm). The tyrosinase inhibitory activity was high with arbutin (95 ppm) and kojic acid (90 ppm). Doenjang-added fermentation materials of the Cudrania tricuspidata fruits were developed, and the organic acid, reducing sugar, and free amino acid of the developed Doenjang were analyzed. The pancreatic lipase inhibitory (PLI) activity and ${\alpha}$-glucosidase inhibitory (AGI) activity of the fermentation materials of the Cudrania tricuspidata fruits and the developed Doenjang were investigated, and it was found that after fermentation, the PLI and AGI activities of the fermentation materials of the Cudrania tricuspidata fruits were higher than those before fermentation, and that the AGI activity of the developed Doenjang after aging ($91.25{\pm}0.04%$) was higher than that before aging ($84.89{\pm}0.08%$).

• Korean Journal of Microbiology
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• v.30 no.4
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• pp.239-245
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• 1992

Buci1llr.s 11c~h~n~1rnSiSi.As 3-2MI which is responsible for the special taste of traditionalKorean soy sause produced two kinds of proteinase. The activity of the proteinasc I washigher about two fold than that of proteinase 11. The optimai, reaction pH of proteinaseI and I1 wcre found to be 7-1 1.5 and 7-9. respectively. Proteinase I1 was more stable andactive than proteinase I at pH ranges around 3 to 5. The optimal te~tlperature of proteinaseI and I1 were 502. The temperature stabilitl of proteinase I1 was Inore stable thanproteinase 1 at temperature range around 30-quot;~A. ctivities of proteinase I and I1 graduallydeclined above $30^{\circ}$C and 45C. respectively. Proteinasc 1 was more active than proteinaseI1 at salt concentration range around 25-3500. The K,,, values of casein and soy proteinfor proteinase I were 6.89 mglml and 3.98 mglml. In case of proteinase 11. they were 9.00mgiml anti 11.44 111g/ml. respectively. The activity of the crudc enzyme was increased by1 rnM Pb(CH3COO). but was decreased by 5 n1M and 10 rnM of HgS04 and ZnS04. Thetwo proteinases produced amino acids and peptides from the soybean protein. The peptideswere digested into amino acids. Both protcinases were found to be the main enzymes thatproduced amino acids which make the main taste of traditional Korean soy sauce.al Korean soy sauce.

• Korean Journal of Microbiology
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• v.53 no.1
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• pp.39-48
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• 2017

In order to evaluate the diversity of microbial population of Korean traditional Deonjang and Gochujang produced in Jeju, Jeonnam, and Jeonbuk province area, microbial communities were analyzed using next generation sequencing. In this result, the dominant bacteria of Deonjang in three area were Bacillus amyloliquefaciens, Tetragenococcus halophilus, and Bacillus was major dominant bacteria in Jeonnam (43.16%) and Jeonbuk (64.54%) area. But in Jeju area, Bacillus was 0.22%, which was significantly different from the other two. Equally, the dominant fungi of Deonjang in 3 area were Candida versatilis. Common fungus in Jeonnam and Jeonbuk area was Candida sp., respectively, 64.22% and 33.68% and Micor sp. was a common fungus in Jeonnam (15.66%) and Jeonbuk area (36.73%). But in Jeju area, Candida sp. and Zygosaccharomyces rouxii were dominant than mold. Bacillus subtilis, Bacillus licheniformis, and B. amyloliquenfaciens were the preminant bacteria in the traditional Gochujang in three regions. But there were no common dominant fungi in the 3 regions. Aspergillus sp. and Rhizopus sp. prevailed in Jeju and Jeonnam region, and Zygosaccharomycess rouxii predominanted in Jeonbuk area. These results suggested that the difference in the samples collected for the study were classified into similar groups according to the characteristics of each sample rather than regional characteristics.

• Journal of Life Science
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• v.33 no.11
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• pp.897-904
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• 2023

This study was done to develope genetic markers with the unique characteristics of genes according to the genomic information of Bacillus velezensis K10. B. velezensis K10 maintained a total of 4,159,835 bps, which was found to encode 5,136 open reading frames (orfs). B. velezensis K10 was found to have much more gene migration due to external factors overall compared to standard strain B. velezensis JS25R. In order to discover genetic selection markers, orfs on the genome to be easily induced to gene mutation were surveyed such as recombinase, integrase, transposase, and phage-related genes. As a result of the investigation, 9 candidate markers were isolated with high possibility as genetic selection markers. Although a part in the various origin's areas showed specificities in comparison with homology, the selected markers were all existed in phage-related areas because they were relatively lower homologies in phage-related genes. PCR analysis was done on B. licheniformis K12, B. velezensis K10, B. subtilis, and B. cereus to establish them as inter-species candidate selection markers. As a result, it was confirmed that B. velezensis K10-specific PCR products were formed in a total of 6 primer sets such as BV3 and BV5 to 9. On the other hand, analysis at the subspecies level observed the formation of B. velezensis K10-specific PCR products in 4 primer sets such as BV3, 5, 8, and 9. Among them, since BV5 and BV8 were detected by very specific results, we suggest that BV5 and 8 can be used as B. velezensis K10 gene selection markers at the species and sub-species level.

• Korean Journal of Food Science and Technology
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• v.40 no.5
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• pp.562-567
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• 2008

The primary objective of this study was to select a probiotic starter for cheonggukjang using 60 strains isolated from rice straw. Among isolated strains, only 8 strains including strain B-59 evidenced proteolytic, amylolytic and soybean activity. These 8 strains were all gram-positive, spore-forming rods. The B-59 strain evidenced antimicrobial activity against Staphylococcus aureus, Listeria monocytogenes, Pseudomonas fluorescens, and Vibrio parahaemolyticus. The antimicrobial activity of isolated B-59 was verified by its ability to inhibit the growth of S. aureus, L. monocytogenes, P. fluorescens, and V. parahaemolyticus. The selected B-59 strain was indentified as Bacillus licheniformis, as shown by a result of 99.0% homology upon API kit analysis. The selected B-59 strain also displayed viability in pH 2.5 artificial gastric juice, artificial bile acid, NaCl (2, 4, 8, 16, 32%), and ethanol (4, 8, 16, 32%). The antioxidative activity of the strain B-59 culture was assessed via a DPPH free radical scavenging activity assay. The activity increased with an increasing in the fermentation time of strain B-59 for 20 hours.