• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.576-584
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• 2017

Objective: To generate recombinant Bacillus subtilis (B. subtilis) engineered for expression of porcine ${\beta}-defensin-2$ (pBD-2) and cecropin P1 (CP1) fusion antimicrobial peptide and investigate their anti-bacterial activity in vitro and their growth-promoting and disease resisting activity in vivo. Methods: The pBD-2 and CP1 fused gene was synthesized using the main codons of B. subtilis and inserted into plasmid pMK4 vector to construct their expression vector. The fusion peptide-expressing B. subtilis was constructed by transformation with the vector. The expressed fusion peptide was detected with Western blot. The antimicrobial activity of the expressed fusion peptide and the recovered pBD-2 and CP1 by enterokinase digestion in vitro was analyzed by the bacterial growth-inhibitory activity assay. To analyze the engineered B. subtilis on growth promotion and disease resistance, the weaned piglets were fed with basic diet supplemented with the recombinant B. subtilis. Then the piglets were challenged by enteropathogenic Escherichia coli (E. coli). The weight gain and diarrhea incidence of piglets were measured after challenge. Results: The recombinant B. subtilis engineered for expression of pBD-2/CP1 fusion peptide was successfully constructed using the main codons of the B. subtilis. Both expressed pBD-2/CP1 fusion peptide and their individual peptides recovered from parental fusion peptide by enterokinase digestion possessed the antimicrobial activities to a variety of the bacteria, including gram-negative bacteria (E. coli, Salmonella typhimurium, and Haemophilus parasuis) and grampositive bacteria (Staphylococcus aureus). Supplementing the engineered B. subtilis to the pig feed could significantly promote the piglet growth and reduced diarrhea incidence of the piglets. Conclusion: The generated B. subtilis strain can efficiently express pBD-2/CP1 fusion antimicrobial peptide, the recovered pBD-2 and CP1 peptides possess potent antimicrobial activities to a variety of bacterial species in vitro. Supplementation of the engineered B. subtilis in pig feed obviously promote piglet growth and resistance to the colibacillosis.

• Food Science and Preservation
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• v.23 no.2
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• pp.233-238
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• 2016

A effect of extraction methods, including stirrer extraction method (SE), ultrasonification extraction method (USE), reflux extraction method (RE), autoclave extraction (AE) and low temperature high pressure extraction (LE) method on the antioxidative and antimicrobial activities of ethanol extracts of Oenothera biennis was investigated. The extraction yield (46.33%), total polyphenol (463.05 mg GAE/g) and flavonoid (71.71 mg RHE/g) content of Oenothera biennis extract obtained by RE were higher than those from other extraction methods. The antimicrobial activity of Oenothera biennis extract was only observed against Bacillus cereus among other tested organisms (Bacillus cereus, Staphylococcus aureus, Escherichia coli and Salmonella Typhimurium). Oenothera biennis obtained by RE showed the best DPPH radical scavenging ability (74.40%), ABTS radical scavenging ability (65.29%), reducing power (1.370 ($OD_{700}$)) and ferrous ion chelating ability (90.14%) compared with other tested extraction methods tested. The RE method was the most efficient method for extracting crude antioxidant and antimicrobial substances from Oenothera biennis. These results suggested that Oenothera biennis obtained by RE could be used as a bioactive and functional material in the food industry.

• Food Science and Preservation
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• v.18 no.1
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• pp.46-52
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• 2011

Wild grape juice was fermented by Gluconacetobacter hansenii TF-2 isolated from tea fungus, to develop a new acidic beverage (fermented wild grape beverage, WGB). Broth was prepared by fermentation of 11~17% (v/v) juice, and sweetened with sucrose (initial sucrose level: $10^{\circ}$ Brix). Fermentation was initiated by addition of 5% (w/v) seed gel (the pellicle of the tea fungus) which had been previously cultured in the same medium (freshjuice broth), and fermentation proceeded in the dark at $29{\pm}1^{\circ}C$ for about 15 days. The major acids produced were succinic acid, malic acid, and acetic acid. After 15 days of fermentation, the organic acid content (principally succinic acid) was 49.6 ppm in WGB 11 and 77.4 ppm in WGB 17. The free sugar content of WGB was 1063.6-1082.5 mg/mL, composed of unfermented fructose, glucose, and sucrose, in that order. The microbial inhibitory effects of the fermented beverage were most apparent when Gram-negative bacteria (Escherichia coli and Salmonella typhimurium) were tested; the inhibition rate was 34.46-88.00%. The new fermented beverage thus displays effective antimicrobial activity against some species of bacteria.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.7-13
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• 2011

This study was conducted to investigate the antimicrobial activity of Ginkgo biloba L. leaves, seed and outer seedcoat against bacteria. Antimicrobial effects of Ginkgo biloba L. leaves (GBL), seed (GBS) and outer seedcoat (GBO) were examined by paper disc method and optical density method to determine minimum inhibition concentration (MIC), and observed by scanning electron microscope (SEM) to figure out the morphological change on the surface when Ginkgo biloba leaves extract was treated. The extracts of GBL, GBS and GBO were extracted by solvents such as methanol, ethanol, water. The methanol extract of GBL and GBO showed the highest antimicrobial activity against Bacillus cereus, Bacillus subtilis, Klebsiella pneumoniae, Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus, Yersinia enterocolitica except Escherichia coli and thus was further fractionated. The MICs of the chloroform fraction of GBL methanol extract were $125{\mu}g$/mL against B. subtilis, and L. monocytogenes; GBO methanol extract were $62.5{\mu}g$/mL against B. cereus and $125{\mu}g$/mL against B. subtilis, and L. monocytogenes. The microorganisms were treated with chloroform extracts ($2000{\mu}g$/mL) of GBL and GBO methanol extracts. It was observed by scanning electron microscope (SEM). The cells were expanded and a part of cell wall was completely destructed by GBL and GBO. Thus Ginkgo biloba L. leaves and outer seedcoat could be further developed into a natural antimicrobial agent.

• Journal of Life Science
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• v.30 no.10
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• pp.896-904
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• 2020

The purpose of this study was to investigate the probiotic characteristics and immune activities of selected lactic acid bacterial (LAB) strains as feed additives in livestock. 301 LAB strains isolated from traditional fermented foods were first assessed for their antibacterial activity potential. Of the 301 isolates, five showed antibacterial activity against five livestock pathogens (Esherichia coli KCCM11234, Listeria monocytogens KCTC3710, Salmonella Typhimurium KCTC1926, Staphylococcus aureus KCCM11593, and Shigella flexneri KCTC2517). The probiotic characteristics of the five selected strains were also investigated by antioxidative activity, hemolysis, bile salt hydrolase, acid resistance and bile tolerance. The SRCM102607 strain was found to have superior probiotic properties and was selected for further experimentation. 16S rRNA gene sequencing showed that SRCM102607 is Pediococcus acidilactici, which was labeled as P. acidilactici SRCM102607 (KCCM 12246P). The survival characteristics of P. acidilactici SRCM102607 in artificial gastrointestinal conditions were assessed under exposed acidic (pH 2.0) and bile (0.5% and 1.0%) conditions. P. acidilactici SRCM102607 was also confirmed to have resistance to various antibiotics, including amikacin, gentamicin, vancomycin, and etc. The TNF-α production by P. acidilactici SRCM102607 was 171.86±4.00 ng/ml. These results show that P. acidilactici RCM102607 has excellent potential for use as a probiotic livestock feed additive.

• Journal of Food Hygiene and Safety
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• v.13 no.4
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• pp.337-343
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• 1998

In order to develop antimicrobial substances, many kinds of medicinal herbs were extracted with absolute ethanol and then antimicrobial activities against various microorganisms were investigated. Ethanol extract from cinnamon bark showed the strongest antimicrobial activity on the growth of almost all submitted microorganisms. Specially, molds such as Aspergillus sp. and Pencillium sp. were inhibited strongly. Therefore, the crude antimicrobial substance from the ethanol extract was fractionated with various solvents such as n-hexane, chloroform, ethyl acetate, and n-butyl alcohol and then their antimicrobial activities were tested. Among the various solvent fractions from the ethanol extract, n-hexane fraction was the best in antimicrobial activity especially against molds. There were no significant changes in antimicrobial activity of the n-hexane fraction by heat treatment at $100^{\circ}C$ for 60 min or $121^{\circ}C$ for 15 min and by the change of pH 4.0~10.0. We could get the results that the n-hexane fraction of cinnamon bark extract showed not only antimutagenicity but also no mutagenicity by Ames test with Salmonella typhimurium TA 98 and TA 100.

• Korean journal of food and cookery science
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• v.17 no.3
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• pp.218-223
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• 2001

This study was conducted to evaluate the functional properties of onion, rosemary and thyme extracts which are the ingredients of onion Kimchi. Onion extract showed a significant difference in antioxidative effect based on peroxide value and thiocyanate method. Antioxidative activity of rosemary extract was similar to those of BHA and BHT. but thyme extract did not show any antioxidative effect.. Onion, rosemary, and thyme extracts showed antimicrobial activities against gram positive bacteria but onion and thyme extracts did not against E. coli, Candida, and molds. Rosemary extract demonstrated a strong activity against L. plantarum which is a major lactobacillus in Kimchi fermentation, and Micrococcus luteus. Onion, rosemary, and thyme extracts showed an effect or hangover relief effect by lowering the alcohol concentration of blood in rat. Blood pressure of the male spontaneous hypertensive rat was suppressed by onion extract after 3 days of feeding, but rosemary and thyme extracts were not effective for lowering blood pressure.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1432-1438
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• 1998

The gamma-radiation sensitivity of eight kinds of pathogenic bacteria related to beef was investigated in frozen cells $(-18^{\circ}C)$ with 0.1 M phosphate buffer and inoculated cells in beef. In frozen cells, D10 values of pathogenic bacteria related to beef were $0.07{\sim}0.69$ kGy, and inactivation factors were $2.90{\sim}42.86$ at the radiation doses of $2{\sim}3$ kGy. Beef was inoculated with Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Enterobacter aerogenes, Vibrio parahaemolyticus, Staphylococcus aureus, Listeria monocytogenes, and Aeromonas hydrophila. Inoculated beef samples were packaged in air and irradiated at 0.005 to 3.0 kGy. Ninety percent of the viable pathogenic bacteria in beef was eliminated by doses of $0.1{\sim}0.61$ kGy at room temperature, and the inactivation factors were $3.28{\sim}30.0$ kGy at the radiation doses of $2{\sim}3$ kGy. Therefore, irradiation is considered to be an effective method to control pathogenic bacteria in beef.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.127-131
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• 2020

Objective: This study was conducted to determine catalytic properties of wheat phytase with exopolyphosphatase activity toward medium-chain and long-chain inorganic polyphosphate (polyP) substrates for comparative purpose. Methods: Exopolyphosphatase assay of wheat phytase toward polyP75 (medium-chain polyP with average 75 phosphate residues) and polyP1150 (long-chain polyP with average 1150 phosphate residues) was performed at pH 5.2 and pH 7.5. Its activity toward these substrates was investigated in the presence of Mg²⁺, Ni²⁺, Co²⁺, Mn²⁺, or ethylenediaminetetraacetic acid (EDTA). Michaelis constant (K_m) and maximum reaction velocity (V_max) were determined from Lineweaver-Burk plot with polyP75 or polyP1150. Monophosphate esterase activity toward p-nitrophenyl phosphate (pNPP) was assayed in the presence of polyP75 or polyP1150. Results: Wheat phytase dephosphorylated polyP75 and polyP1150 at pH 7.5 more effectively than that at pH 5.2. Its exopolyphosphatase activity toward polyP75 at pH 5.2 was 1.4-fold higher than that toward polyP1150 whereas its activity toward polyP75 at pH 7.5 was 1.4-fold lower than that toward polyP1150. Regarding enzyme kinetics, K_m for polyP75 was 1.4-fold lower than that for polyP1150 while V_max for polyP1150 was 2-fold higher than that for polyP75. The presence of Mg²⁺, Ni²⁺, Co²⁺, Mn²⁺, or EDTA (1 or 5 mM) exhibited no inhibitory effect on its activity toward polyP75. Its activity toward polyP1150 was inhibited by 1 mM of Ni²⁺ or Co²⁺ and 5 mM of Ni²⁺, Co²⁺, or Mg²⁺. Ni²⁺ inhibited its activity toward polyP1150 the most strongly among tested additives. Both polyP75 and polyP1150 inhibited the monophosphate esterase activity of wheat phytase toward pNPP in a dose-dependent manner. Conclusion: Wheat phytase with an unexpected exopolyphosphatase activity has potential as a therapeutic tool and a next-generational feed additive for controlling long-chain polyP-induced inappropriate inflammation from Campylobacter jejuni and Salmonella typhimurium infection in public health and animal husbandry.

• KSBB Journal
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• v.23 no.2
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• pp.147-152
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• 2008

The variety of functional plants has an attention for new natural food preservation and natural antiseptic development. The extracts from functional plants with various methods (ethanol extraction, hot water extraction and supercritical fluid extraction) tested antimicrobial activity against 10 strains including the pathogenic and food poisoning bacteria, the yeast and fungi. The antimicrobial activities of supercritical fluid extracts were shown higher than ethanol extract and hot water extract when tested with disc-diffusion method and minimum inhibitory concentration (MIC). Antimicrobial activity of supercritical fluid extract was two times higher than ethanol extract in Cinnaonomum cassia. In addition, the supercritical fluid extractions of Chamaecyparis obtuas and the C. cassia showed the higher yield than Origanum vulgare and Scutellariae baicalensis. The supercritical fluid extract of C. cassia showed an antimicrobial activity against all strains tested. The supercritical fluid extract of S. baicalensis showed strong antimicrobial activity on Listeria monocytogenes. Supercritical fluid extraction of O. vulgare and C. obtuas showed strong antimicrobial activity on Salmonella typhimuriium. In MIC test, C. obtuas was shown the best natural material for the preparation of natural antimicrobial agent by supercritical fluid extraction. In conclusion, these results suggest that supercritical fluid extraction technique was effective to obtain functional ingredient with higher antimicrobial activity in the development of new antimicrobial reagent from natural materials.