• 한국축산식품학회지
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• 제33권5호
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• pp.595-602
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• 2013

As a major inhibitory neurotransmitter of the central nervous system in animals, ${\gamma}$-aminobutyric acid (GABA) has several physiological functions, such as anti-hypertensive, diuretic, tranquilizer and anti-stress effects in human. In order to determine strains with high GABA producing ability and glutamate decarboxylase (GAD) activity, 273 bacteria were isolated from various types of Kimchi. Strain K255 contained $386.37{\mu}g/mL$ of GABA in MRS broth containing 1% MSG, $600.63{\mu}g/mL$ of GABA in MRS broth containing 2% MSG and $821.24{\mu}g/mL$ of GABA in MRS broth containing 3% MSG. It showed that K255 had the highest GABA production ability compared to other commercial lactic acid bacteria. K255 was identified as Lactobacillus plantarum based on its API carbohydrate fermentation pattern and 16S rDNA sequence. K255 was investigated for its physiological characteristics. The optimum growth temperature of K255 was $37^{\circ}C$and cultures took 13 h to reach the pH 4.4. K255 showed more sensitive to bacitracin in a comparison of fifteen different antibiotics, and showed most resistance to kanamycin and vancomycin. Moreover, it was comparatively tolerant to bile juice and acid and displayed resistance to Escherichia coli, Salmonella Typhimurium, Staphylococcus aureus, with rates of 30.8%, 29.7%, and 23.4% respectively. These results demonstrate that K255 could be an excellent strain for the production of functional products.

• 한국축산식품학회지
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• 제40권4호
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• pp.512-526
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• 2020

Synthetic nitrite is considered an undesirable preservative for meat products; thus, controlling synthetic nitrite concentrations is important from the standpoint of food safety. We investigated 1,000 species of microorganisms from various kimchi preparations for their potential use as a starter culture for the production of nitrites. We used 16S rRNA gene sequence analysis to select a starter culture with excellent nitrite and nitric oxide productivity, which we subsequently identified as Staphylococcus hominis subspecies hominis WiKim0113. That starter culture was grown in NaCl (up to 9%; w/v) at 10℃-40℃; its optimum growth was observed at 30℃ at pH 4.0-10.0. It exhibited nonproteolytic activity and antibacterial activity against Clostridium perfringens, a bacterium that causes food poisoning symptoms. Analysis of Staphylococcus hominis subspecies hominis WiKim0113 with an API ZYM system did not reveal the presence of β-glucuronidase, and tests of the starter culture on 5% (v/v) sheep blood agar showed no hemolytic activity. Our results demonstrated the remarkable stability of coagulase-negative Staphylococcus hominis subspecies hominis WiKim0113, especially in strain negative for staphylococcal enterotoxins and sensitive to clinically relevant antibiotics. Moreover, Staphylococcus hominis subspecies hominis WiKim0113 exhibited a 45.5% conversion rate of nitrate to nitrite, with nitrate levels reduced to 25% after 36 h of culturing in the minimal medium supplemented with nitrate (200 ppm). The results clearly demonstrated the safety and utility of Staphylococcus hominis subspecies hominis WiKim0113, and therefore its suitability as a starter culture.

• 한국미생물·생명공학회지
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• 제37권1호
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• pp.24-32
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• 2009

숙성 정도가 오래된 김장 배추 김치에서 분리되어 16S rDNA 염기 서열 분석을 통해 부분 동정된 KNUC25 분리 균주를 단백질 전기 영동 패턴과 생리적 특징 그리고 염기 서열의 유사성을 비교하여 Lactobacillus paraplantarum로 동정하였다. L. paraplantarum KNUC25의 농축 상등액은 그람 양성균과 음성균에 넓은 범위의 항균 활성을 나타냈다. 전자 현미경을 통해 KNUC25가 만들어내는 항균 물질은 세균의 표면에 작용하여 생육을 억제하는 것으로 확인되었다. 항균 활성 물질을 생산하는 최적 온도는 섭씨 30도이고, 높은 온도에서도 항균 활성을 보였으며 단백질 분해 효소에 안정하며 비단백질성 항균 물질일 가능성을 보여주었다.

• Journal of Dairy Science and Biotechnology
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• 제24권1호
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• pp.19-28
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• 2006

유아 분변으로부터 분리한 52 균주는 그람 양성균인 Listeria monocytogenes KCCM 40307$^T$에 대하여 항균활성이 있었고 그 중에서도 선별 균주 A24의 항균활성이 45% 이상으로 가장 높았다. Bifidobacterium longum A24의 생육 및 항균 물질 생산을 검토하였을 때 균체의 생육은 28시간 배양 시 최고에 도달하였고 항균 활성은 36시 간 배양 시 최고를 나타내었다. 선별균주 A24는 16S rRNA-based molecular typing 결과Bifidobacterium 속 균주임을 확인할 수 있었고 형태학적 ${\cdot}$ 생화학적인 방법으로 검토하여 보았을 때 Bifidobacterium longum으로 판단되었으며, 16s rDNA sequencing 결과 최종적으로 Bifidobacterium longum로 동정 되었으며 이것을 Bifidobacterium longum A24로 명명하였다. Bifidobacterium longum A24의 항균 활성 물질은 균체의 생육이 가장 좋은 28시간 배양에서가 아니라 그보다 늦은 36시간 배양에서 최고를 나타내었고 그 이후로는 활성이 감소하는 경향을 보였다. 이것은 Bifidobacterium longum A24이 생성하는 항균 활성 물질이 bacteriocin과 같은 2차 대사 산물임을 암시하는 결과로 해석된다.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1688-1694
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• 2009

A novel strain of fluorescent pseudomonad (PB-St2) was isolated from surface-sterilized stems of sugarcane grown in Pakistan. The bacterium was identified as Pseudomonas aurantiaca on the basis of 16S rRNA gene sequence analysis and results from physiological and biochemical characteristics carried out with API50 CH and QTS 24 bacterial identification kits. Assays using substrate-specific media for enzymes revealed lipase and protease activities but cellulase, chitinase, or pectinase were not detected. The bacterium was unable to solubilize phosphate or produce indole acetic acid. However, it did produce HCN, siderophores, and homoserine lactones. In dual culture assays on agar, the bacterium showed antifungal activity against an important pathogen of sugarcane in Pakistan, namely Colletotrichum falcatum, as well as for pathogenic isolates of Fusarium oxysporium and F. lateritium but not against F. solani. The antifungal metabolites were identified using thin-layer chromatography, UV spectra, and MALDI-TOFF spectra and shown to be phenazine-1-carboxylic acid (PCA), 2-hydroxyphenazine (2-OH-PHZ), and N-hexanoyl homoserine lactone (HHL) (assessed using only TLC data). The capacity of this bacterium to produce HCN and 2-OH-PHZ, as well as to inhibit the growth of C. falcatum, has not been previously reported.

• 한국식품영양학회지
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• 제27권2호
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• pp.213-218
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• 2014

적리를 동반하는 돼지 분변에서 용혈성 균주를 분리 동정한 후, 장 독소 생성 유 무 및 항생제 감수성 시험을 수행하였다. 분리된 용혈성 균주 B. cereus BY06의 gyrB 유전자 염기서열을 분석한 결과, B. cereus와 99% 유사성을 나타내었다. PCR법에 의한 장 독소 유전자 검출 시험에서 B. cereus BY06는 장 독소 분비 양성으로 판정됨에 따라 설사형 식중독 균임이 확인되었다. B. cereus BY06를 이용한 항균제의 감수성 시험 결과, penicillin G에는 내성을 나타낸 반면 cephalothin, vancomycin, clindamycin, fusidic acid, gentamicin, ciprofloxacin, tetracycline 및 rifampin에 대하여 감수성을 나타냈다. 본 연구를 통해 돼지 분변에서 분리된 B. cereus 균주는 설사를 유발하는 장 독소를 분비하며, penicillin G에 대한 내성을 확인하였다.

• 한국축산식품학회지
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• 제30권5호
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• pp.804-810
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• 2010

This study was performed to select protease-producing Bacillus sp. as a potential probiotic starter for fermented meat products. In order to isolate protease-producing bacterium from meju, measured the diameter of the clear zone on agar plate (TSA, 1% (w/v) skim milk) and analyzed for intracellular protease activity, then 10 Bacillus-like strains were isolated. Three Bacillus-like strains (P19, P27, and P33) among 10 strains were able to tolerate in acidic condition (TSB, pH 2.5, 2 h incubation). These 3 strains were showed antimicrobial activity against food-borne pathogenic bacteria. These vegetative cells of 3 strains were showed a survival rate of 0.04% to 0.08% under the artificial gastric acidic condition (TSB, pH 2.5 with 1% (w/v) pepsin), but spore-forming cells were 56.29% to 84.77%. Vegetative cells of 3 strains were the least bile-resistant, while spore-forming cells of 3 strains showed higher survival rate more than 76% under artificial bile condition (TSB, 0.1% (w/v) oxgall bile). In these strains, P27 strain was finally selected as a good probiotic strain. P27 strain was tentatively identified as Bacillus amyloliquefaciens by API CHB kit and 16S rDNA sequence analysis. The results of this study suggest that B. amyloliquefaciens P27 can be used as a potential probiotic starter for fermented meat product.

• 한국축산식품학회지
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• 제32권5호
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• pp.635-643
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• 2012

As an appraisal for the application of a new starter culture, more than 200 lactic acid bacteria strains were isolated from raw milk and healthy human feces. The strains showing excellent growth and acid production ability in 10% skim milk media were selected and identified as Lactobacillus casei based on the results of their API carbohydrate fermentation patterns, as well as 16S rDNA sequence analysis. To assess the effect of L. casei strains on irritable bowel disease (IBD), the inhibitory effect of the selected strains against the nitric oxide (NO) production of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells was measured. Among the tested L. casei strains, L. casei MCL was observed to have the greatest NO inhibitory activity. Additionally, L. casei MCL was found to inhibit mRNA expression of pro-inflammatory cytokines (interleukin-$1{\beta}$, IL-6, TNF-${\alpha}$), as well as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) involved in pathophysiologic processes such as inflammation. The mRNA expression of anti-inflammatory cytokines, including IL-10 and transforming growth factor-$1{\beta}$ (TGF-${\beta}$) of L. casei MCL, was confirmed using quantitative real-time PCR. In conclusion, L. casei MCL showed decreases in the expression of pro-inflammatory cytokines and up-regulated expression of the anti-inflammatory cytokine.

• Food Science and Biotechnology
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• 제18권5호
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• pp.1100-1108
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• 2009

Lactic acid bacteria (LAB) isolated from various fruits and vegetables were screened in order to determine appropriate fermentation starters for manufacturing functional fermented onion juice. From the initial screening test comprising more than 700 isolated LAB, 16 isolates were selected based on their acid production rate. Among the selected isolates, the fermentation broth of KC-007 exhibited the highest electron donating and nitrite scavenging activities, with values at pH 1.2 of 95.6 and 68.7%, respectively. From the overall results obtained in this study, we finally selected the bacterium KC-007 as a fermentation starter. This bacterium was identified and named as Pediococcus pentosaceus based on its morphological and physiological characteristics, carbon-utilization pattern (as assessed using an API 50CHL kit), and molecular genetic characteristics (as assessed using the nucleotide sequence of the 16S rRNA gene). The optimal temperature, pH, and starter inoculation concentration (v/v) required for growth of the isolated strain were $40^{\circ}C$, pH 4.0-6.0, and 2%(v/v), respectively.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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• pp.21-35
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• 1999

We have previously isolated OsMADS4 gene that is a member of the class B MADS box genes from rice. In this study, another member of the class B MADS box genes was isolated from rice flower by the yeast two-hybrid screening method using OsMADS4 as bait. RNA blot analyses revealed that the clone, OsMADS16, was expressed in the second and third whorls, whereas the OsMADS4 transcripts were present in the second, third, and fourth whorls. These expression patterns of the OsMADS16 and OsMADS4 genes are very similar with those of AP3 and PI, the class B genes of Arabidopsis, respectively. In the yeast two-hybrid system, OsMADS4 interacted only with OsMADS16 among several rice MADS genes investigated, suggesting that OsMADS4 and OsMADS16 function as a heterodimer in specifying sepal and petal identities. We have also isolated OsMADS6 gene using OsMADS1 as a probe. Both are members of the AGL2 MADS family. Various MADS genes that encode for protein-protein interaction partners of the OsMADS6 protein were isolated by the yeast two-hybrid screening method. A majority of these genes belong to the AGL2 family. Sequence Homology, expression pattern, and ectopic expression phenotypes indicated that one of the interaction partners, OsMADS14, appears to be homologous to API, the class A MADS gene of Arabidopsis.