• Korean Journal of Food Science and Technology
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• v.48 no.4
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• pp.306-316
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• 2016

This study was conducted to evaluate the functional characteristics and safety properties of lactic acid bacteria isolated from salt-fermented anchovy, a putative probiotic candidate. The following isolates were identified by biochemical profiles, carbohydrate fermentation patterns, and 16S rRNA sequencing: Enterococcus faecium AJ06, Leuconostoc mesenteroides AJ13, Pediococcus halophilus AJ22, Lactobacillus sakei AJ29, and Pediococcus pentosaceus AJ35. The strains AJ06, AJ22, AJ29 exhibited high tolerance to simulated gastric and intestinal juices and were able to produce bile salt hydrolase on MRS agar plates supplemented with taurocholic acid and/or taurodeoxycholic acid. The strains AJ22 and AJ29, which demonstrated high adherence to Caco-2 cells and resistance to various antibiotics, effectively inhibited the growth of food-borne pathogens by the production of antimicrobial substances. These strains did not show ${\alpha}-$ or ${\beta}$-haemolysis on blood agar. Furthermore, biogenic amines in MRS broth containing the precursor amino acids were not mutagenic in Salmonella Typhimurium TA98 and TA100.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.78-83
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• 2004

In order to screen microorganisms producing phopholipase D (PLD) had high transphosphatidylation activity, about 1,000 Actinomycetes strains were isolated from the 63 soil samples, collected over 6 local area in Korea. When the hydrolytic activity in the supernatant was determined, 131 strains produced PLD more than 0.3U/$m\ell$. Among 131 culture broths tested, 23 ones had transphosphatidylation activity higher than 20% and finally one strain (Actinomycetes KF923), which had highest hydrolytic and transphophadylation activity, was selected. Actinomycetes KF923 showed the highest hydrolytic activity (13U/$m\ell$) and phosphatidylation activity (95%) after 48 h fermentation using the P medium (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, $CaCO_3$ 0.4%, pH 7.2). PLD was purified from the culture broth of Actinomycetes KF923 and the specific activity of purified PLD was 567U/mg. The molecular weight of PLD was about 55kD and the optimum pH and temperature were pH 6.0 and $60^{\circ}C$, respectively. The stability of PLD toward pH and temperature were high around pH 8.0 and below $40^{\circ}C$ Special metal ions were not necessary to the PLD activity.

• Microbiology and Biotechnology Letters
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• v.9 no.4
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• pp.197-205
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• 1981

Penicillin amidase of Bacillus megaterium was recovered from the fermentation broth by adsorption on celite and immobilized by entrapping the adsorbed enzyme in acrylamide gel. The operational stability in column reactor was greatly increased by entrappment as compared with that of without entrappment. The optimum pH of the immobilized enzyme was 8.7 with broader activity profile than that of the free enzyme, while the most stable pH range appeared to be between pH 7.5 and 8.0. The optimum temperature was shifted to 5$0^{\circ}C$ from 45$^{\circ}C$ for the soluble enzyme. The values of Km and the inhibition constants for 6-APA( $K_{ia}$ ) and phenylacetic acid ( $K_{ip}$ ), were 4.55 mM, 36.5mM, and 10.5mM, respectively. No significant internal pore diffusion limitation was found since the value of effectiveness factor was 0.95. The operational half life in a column reactor at pH 8.0 was 6.8 days at 4$0^{\circ}C$ and 47 days at 3$0^{\circ}C$, whereas that of without entrappment was only 1 day and 4 days, respectively. The performance of a batch and a column reactor was also discussed with respect to the productivity. The results demonstrated that the entrappment of an adsorbed enzyme for the enhancement of the operational stability of the immobilized enzyme was useful especially when an extracellular enzyme was used.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.21 no.2
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• pp.71-79
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• 1988

For the purpose of producing low salt fermented anchovy by accelerated method with a strong proteolytic bacteria, in this study, a strong proteolytic bacterium was isolated from low salt fermented anchovy and its bacteriological characteristics and properties of protease were experimented. The results obtained were as fellows : three proteolytic bacteria, Aeromonas anaerogenes Barillus subtilis and Staphylococcus saprophyticus were isolated from low salt fermented anchovy($4\%\;of\;salt,\;4\%\;of\;KCl,\;0.5\%\;of\;lactic\;acid,\;6\%$of sorbitol and $4\%$ of alcohol extract of red pepper) after 40 days fermentation. Among these strains, which grow best at $30^{\circ}C$, pH 7.0, B. subtilis was found the best proteolytic strain and benefit for industrial use as shown $0.95\;hr^{-1}$ of specific growth rate, $89{\mu}g-Tyr/hr.ml$ of maximum activity after 12 hrs culture in TPY broth. The protease produced by by B. subtilis showed maximum activity at $35^{\circ}C$, pH 7.0, and molecular weight was estimated to be 23,000 by Sephadex G-100 filtration, and it was supposed to be a kind of metal chelator sensitive neutral protease from the results of strong sensitivity against EDTA, o-phenanthroline and metal ions such as $Cu^{2+},\;Ni^{2+},\;Fe^{2+}.Km$ value of that by method of Lineweaver-Burk was determinded to be $0.73\%$ for casein as a substrate.

• The Plant Pathology Journal
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• v.35 no.6
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• pp.662-673
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• 2019

Root-knot nematodes (RKNs) are among the most destructive plant-parasites worldwide, and RKN control has been attempted mainly using chemical nematicides. However, these chemical nematicides have negative effects on humans and the environment, thus necessitating the search for eco-friendly alternative RKN control methods. Here, we screened nematicidal lactic acid bacteria (LAB) isolated from kimchi and evaluated their efficacy as biocontrol agents against RKNs. Of 237 bacterial strains, Lactobacillus brevis WiKim0069 showed the strongest nematicidal activity against the second-stage juveniles (J2) of Meloidogyne incognita, M. arenaria, and M. hapla and inhibited the egg hatch of M. incognita. The culture filtrate of WiKim0069 had a pH of 4.2 and contained acetic acid (11,190 ㎍/ml), lactic acid (7,790 ㎍/ml), malic acid (470 ㎍/ml), and succinic acid (660 ㎍/ml). An artificial mixture of the four organic acids produced by WiKim0069 also induced 98% M. incognita J2 mortality at a concentration of 1.25%, indicating that its nematicidal activity was derived mainly from the four organic acids. Application of WiKim0069 culture filtrate suppressed the formation of galls and egg masses on tomato roots by M. incognita in a dose-dependent manner in a pot experiment. The fermentation broth of WiKim0069 also reduced gall formation on melon under field conditions, with a higher efficacy (62.8%) than that of fosthiazate (32.8%). This study is the first report to identify the effectiveness of kimchi LAB against RKNs and to demonstrate that the organic acids produced by LAB can be used for the RKN management.

• The Plant Pathology Journal
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• v.33 no.3
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• pp.337-344
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• 2017

To develop a commercial product using the mycoparasitic fungus Simplicillium lamellicola BCP, the scale-up of conidia production from a 5-l jar to a 5,000-l pilot bioreactor, optimization of the freeze-drying of the fermentation broth, and preparation of a wettable powder-type formulation were performed. Then, its disease control efficacy was evaluated against gray mold diseases of tomato and ginseng plants in field conditions. The final conidial yields of S. lamellicola BCP were $3.3{\times}10^9conidia/ml$ for a 5-l jar, $3.5{\times}10^9conidia/ml$ for a 500-l pilot vessel, and $3.1{\times}10^9conidia/ml$ for a 5,000-l pilot bioreactor. The conidial yield in the 5,000-l pilot bioreactor was comparable to that in the 5-l jar and 500-l pilot vessel. On the other hand, the highest conidial viability of 86% was obtained by the freeze-drying method using an additive combination of lactose, trehalose, soybean meal, and glycerin. Using the freeze-dried sample, a wettable powder-type formulation (active ingredient 10%; BCP-WP10) was prepared. A conidial viability of more than 50% was maintained in BCP-WP10 until 22 weeks for storage at $40^{\circ}C$. BCP-WP10 effectively suppressed the development of gray mold disease on tomato with control efficacies of 64.7% and 82.6% at 500- and 250-fold dilutions, respectively. It also reduced the incidence of gray mold on ginseng by 65.6% and 81.3% at 500- and 250-fold dilutions, respectively. The results indicated that the new microbial fungicide BCP-WP10 can be used widely to control gray mold diseases of various crops including tomato and ginseng.

• KSBB Journal
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• v.11 no.1
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• pp.15-21
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• 1996

Purification of hyaluronic acid produced by Streptococcus equi was carried out to obtain clinical grade hyaluronic acid. The removal method of the bacteria was selected as filtration because filtration was the most effective method in removing impurities such as protein and nucleic acid of the fermentation broth. The removal efficiencies of protein and nucleic acid of hyaluronic acid solution were increased to 75% and 67%, respectively, by filtration with adding 0.6% of activatied carbon and 1.0% colite. Hyaluronic acid solution was precipitated by mixing with 2 volumes of ethanol. Effects of pH and conductivity on ethanol preciptation of hyaluronic acid were investigated. Protein and nucleic acid of hyaluronic acid were remained almost constant regardless of pH and conductivity, and the recovery of hyaluronic acid was optimum as about 85% at pH 7 and l00mS of conductivity Protein of hyaluronic acid was completly removed by three serial filtration and ethanol precipitation, however, nucleic acid was not removed. Hyaluronic acid solution was passed through a column of Duolite A7 to remove its nucleic acid, where 65% of nucleic acid was removed at pH 7 and 40mS of conductivity. The residual nucleic acid of hyaluronic acid solution was completly removed by treatment of 0.2% hydroxyapatite and the clinical grade hylauronic acid could be obtained.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.12
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• pp.110-116
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• 2019

To investigate the antagonistic inhibitory effects in a mixed culture between probiotics and various pathogenic microorganisms, 140 probiotics were identified using a 16 rRNA sequencing phylogenetic analysis method, and various probiotics strains were isolated from Korean kimchi from January to December 2016. The antagonistic inhibition test of a mixed culture of four probiotics (Enterococcus faecalis, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus reuteri) with excellent antimicrobial activity and six pathogenic microorganisms (Candida albicans, Salmonella Enteritidis, E. coli O157:H7, Shigella flexneri, Staphylococcus aureus, and Pseudomonas aeruginosa)showed that the growth of most probiotics strains increased normally after culture, but growth was inhibited almost completely in most pathogenic microorganisms, except for S. Enteritidis. This antagonistic inhibitory effect in vitro was attributed to the low pH of the lactic acid and organic acid produced during fermentation. As a result, four probiotics strains isolated from Korean Kimchi are very likely to be developed as therapeutic agents for female yeast infections and colon and skin care. In the future, these therapeutic agents will help improve public health related to probiotics.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.119-127
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• 2013

Twenty-three strains of Leuconostoc species and 45 strains of Weissella species inhibiting the growth of Lactobacillus sakei, one of the most populous lactic acid bacteria in over-ripened kimchi, were isolated from kimchi in our previous study. Among these hetero-fermentative 68 strains, Leuconostoc mesenteroides CK0128, Weissella cibaria CK0633, and W. cibaria KK0797 exhibited a relatively high survival rate in MRS medium, which was adjusted to pH 4.3 using an acid mixture consisting of acetic and lactic acids, and produced a large amount of exopolysaccharides. The culture supernatants of 3 strains were fractionated by a molecular weight cutter and lyophilized. The fractions with a molecular weight smaller than 3,000 Da showed antagonistic activity against Staphylococcus aureus and Lb. sakei. The anti-bacterial substances were very stable to heat treatments ($121^{\circ}C$, 15 min) and active at acidic conditions below pH 5. ${\alpha}$-Amylase, lipase, and proteolytic enzymes (proteinase K and pepsin) did not affect their activities. These non-proteinaceous anti-bacterial substances inhibited the growth of several food pathogens.

• Journal of Microbiology and Biotechnology
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• v.14 no.2
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• pp.276-283
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• 2004

Screening was performed to isolate cellulose-producing microorganisms from the Korean traditional fermented persimmon vinegar. The resulting strain, KJ $145^{T}$, was then taxonomically investigated by phenotypic characterization, particularly chemotaxonomic, and by phylogenetic inference based on a 16S rDNA sequence analysis including other related taxa. Strain KJ $145^{T}$ was found to grow rapidly and form pale white colonies with smooth to rough surfaces on a GYC agar. Strain KJ $145^T$ also produced acetate from ethanol, and was tolerable to 10% ethanol in SM medium. In a static culture, a thick cellulose pellicle was produced, and in GYC broth, the strain grew at temperatures ranging from 28 to $40^\circ{C}$ with an optimum pH of 4.0. The genomic DNA G+C content of strain KJ $145^T$ was 61.9 mol%, and the predominant ubiquinone was Q 10 as the major quinone and Q9 as the minor quinone. The major cellular fatty acids were $C_{16:0}$ and the sum in feature 7 ($C_{18:1}$ w9c, w12t and/or w7c). A 16S rRNA-targeted oligonucleotide probe specific for strain KJ $145^T$was constructed, and the phylogenetic position of the new species was derived from a 16S rDNA-based tree. When comparing the 16S rDNA nucleotide sequences, strain KJ $145^T$ was found to be most closely related to G. hansenii LMG $1527^T$ (99.2%), although KJ $145^T$ was still distinct from G. hansenii LMG $l527^T$ and G. xylinus LMG $1515^T$ in certain phenotypic characteristics. Therefore, on the basis of 16S rDNA sequences and taxonomic characteristics, it is proposed that strain KJ $145^T$ should be placed in the genus Gluconacetobacter as a new species, Gluconacetobacter persimmonis sp. nov., under the type-strain KJ $145^T$ (=KCTC =$10175BP^T$=KCCM=$10354^T$).