• Journal of Microbiology and Biotechnology
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• 제21권6호
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• pp.574-581
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• 2011

A Gram-positive bacterium was isolated from the saline soils of Jangpura (U.P.), India, and showed high-level of radiation-resistant property and survived upto 12.5 kGy dose of gamma radiation. The 16S rDNA sequence of this strain was examined, identified as Bacillus sp. strain HKG 112, and was submitted to the NCBI GenBank (Accession No. GQ925432). The mechanism of radiation resistance and gene level expression were examined by proteomic analysis of whole-cell extract. Two proteins, 38 kDa and 86.5 kDa excised from SDS-PAGE, which showed more significant changes after radiation exposure, were identified by MALDI-TOF as being flagellin and S-layer protein, respectively. Twenty selected 2-DE protein spots from the crude extracts of Bacillus sp. HKG 112, excised from 2- DE, were identified by liquid chromatography mass spectrometry (LC-MS) out of which 16 spots showed significant changes after radiation exposure and might be responsible for the radiation resistance property. Our results suggest that the different responses of some genes under radiation for the expression of radiation-dependent proteins could contribute to a physiological advantage and would be a significant initial step towards a fullsystem understanding of the radiation stress protection mechanisms of bacteria in different environments.

• Asian-Australasian Journal of Animal Sciences
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• 제27권5호
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• pp.648-657
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• 2014

Silage making has become a significant method of forage conservation worldwide. To determine how tomato pomace (TP) may be used effectively as animal feed, it was ensilaged for 90 days and microbiology counts, fermentation characteristics and chemical composition of tomato pomace silage (TPS) were evaluated at the 30th, 60th, and 90th days, respectively. In addition, 103 lactic acid bacteria were isolated from TPS. Based on the phenotypic and chemotaxonomic characteristics, 16S rDNA sequence and carbohydrate fermentation tests, the isolates were identified as 17 species namely: Lactobacillus coryniformis subsp. torquens (0.97%), Lactobacillus pontis (0.97%), Lactobacillus hilgardii (0.97%), Lactobacillus pantheris (0.97%), Lactobacillus amylovorus (1.9%), Lactobacillus panis (1.9%), Lactobacillus vaginalis (1.9%), Lactobacillus rapi (1.9%), Lactobacillus buchneri (2.9%), Lactobacillus parafarraginis (2.9%), Lactobacillus helveticus (3.9%), Lactobacillus camelliae (3.9%), Lactobacillus fermentum (5.8%), Lactobacillus manihotivorans (6.8%), Lactobacillus plantarum (10.7%), Lactobacillus harbinensis (16.5%) and Lactobacillus paracasei subsp. paracasei (35.0%). This study has shown that TP can be well preserved for 90 days by ensilaging and that TPS is not only rich in essential nutrients, but that physiological and biochemical properties of the isolates could provide a platform for future design of lactic acid bacteria (LAB) inoculants aimed at improving the fermentation quality of silage.

• Journal of Applied Biological Chemistry
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• 제59권3호
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• pp.173-178
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• 2016

Phenyllactic acid (PLA), which is a known antimicrobial compound, can be synthesized through the reduction of phenylpyruvic acid (PPA) by lactate dehydrogenase of lactic acid bacteria (LAB). PLA-producing LAB was isolated from coffee beans, and the isolated LAB was identified as Lactobacillus zeae Y44 by 16S rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. zeae Y44 was assessed for both its capability to produce the antimicrobial compound PLA and its antifungal activity against three fungal pathogens (Rhizoctonia solani, Botrytis cinerea, and Colletotrichum aculatum). PLA concentration was found to be 4.21 mM in CFS when L. zeae Y44 was grown in MRS broth containing 5 mM PPA for 12 h. PLA production could be promoted by the supplementation with PPA and phenylalanine (Phe) in the MRS broth, but not affected by 4-hydroxy-phenylpyruvic acid, and inhibited by tyrosine as precursors. Antifungal activity assessment demonstrated that all fungal pathogens were sensitive to 5 % CFS (v/v) of L. zeae Y44 with average growth inhibitions ranging from 27.8 to 50.0 % (p<0.005), in which R. solani was the most sensitive with an inhibition of 50.0 %, followed by B. cinerea and C. aculatum. However, pH modification of CFS to pH 6.5 caused an extreme reduction in their antifungal activity. These results may indicate that the antifungal activity of CFS was caused by acidic compounds like PLA or organic acids rather than proteins or peptides molecules.

• 한국미생물·생명공학회지
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• 제44권2호
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• pp.163-170
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• 2016

전국 각지에서 수집된 발표식품으로부터 젖산균을 분리하고 유해균인 B. cereus, L. monocytogenes, S. Typhimurium, S. aureus 및 S. epidermidis 균주에 대하여 생육 억제효능이 강한 L2167 균주를 선발하고, 16S rDNA 분석법을 이용하여 L. plantarum로 동정하였다. L2167 균주의 단백질 분획물을 제작하여 S. aureus 등 5종의 유해균 배양액에 첨가한 후, 전자현미경으로 관찰하였다. L. plantarum L2167 균주의 균체 단백질 분획물은 유해균의 생육을 억제하였으며, 생육 억제활성은 균체 내부에 함유된 단백질이 유해균의 세포막을 파괴하는 것에서 기인하는 것으로 추정되었다. L. plantarum L2167은 pH 5.5로 조절한 MRS 배지에서 35℃로 배양하였을 때, 가장 우수한 비성장속도를 나타냈으며, 대조구로 사용된 L. plantarum KCTC21004보다 NaCl에 대하여 민감한 특징을 나타내었다. 본 연구를 통하여 선발한 L. plantarum L2167 균주는 추가연구를 통하여 유해균의 생육을 효율적으로 억제할 수 있는 스타터 균주로서 개발될 수 있을 것으로 기대된다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.95.2-96
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• 2003

In order to develop a new microbial fungicide for the control of vegetable diseases using endophytic bacteria, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth media, their antifungal activities were screened by in vivo bioassays against Botrytis cinerea(tomato gray mold), Pythium ultimum(cucumber damping-off), Phytopkhora infestans(tomato late blight), Colletotrichum orbiculare(cucumber anthracnose), and Blumeria graminis f. sp. hordei(barley powdery mildew). As the results of screening, 38 bacterial strains showed potent antifungal activities against at least one of 5 plant pathogens. A bacterial strain EB072 displayed potent disease control activities against 3 plant diseases. Among the bacterial strains with a potent antifungal activity against cucunlber anthracnose, three bacterial strains, EB054, EB151 and EB215, also displayed a potent in vitro antifungal activity against C. acutatum, a fungal agent causing pepper anthracnose. A bacterial strain EB215 obtained from roots of cucumber was identified as Burkholderia cepacia based on its physiological and biochemical characteristics and 165 rRNA gene sequence. An antifungal substance was isolated from the liquid cultures of B. cepacia EB215 strain by ethyl acetate partitioning, repeated silica gel column chromatography, and invitro bioassay, Its structural determination is in progress by various instrumental analyses.

• The Plant Pathology Journal
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• 제22권1호
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• pp.36-45
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• 2006

Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

• 한국버섯학회지
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• 제14권4호
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• pp.162-167
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• 2016

Lactic acid bacteria (LAB) producing phenyllactic acid (PLA), which is known as antimicrobial compound, was isolated from button mushroom bed and the isolated LAB was identified to Lactobacillus casei by 16 rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. casei was assessed for both the capability to produce the antimicrobial compound PLA and the antifungal activity against three fungal pathogens (Rhizoctonia solani, Botrytis cinerea, and Collectotricum aculatum). PLA concentration was investigated to be 3.23 mM in CFS when L. casei was grown in MRS broth containing 5 mM phenylpyruvic acid as precursor for 16 h. Antifungal activity demonstrated that all fungal pathogens were sensitive to 5% CFS (v/v) of L. casei with average growth inhibitions ranging from 34.58% to 65.15% (p < 0.005), in which R. solani was the most sensitive to 65.15% and followed by C. aculatum, and B. cinerea. The minimum inhibitory concentration (MIC) for commercial PLA was also investigated to show the same trend in the range of 0.35 mg mL-1 (2.11 mM) to 0.7 mg mL-1 (4.21 mM) at pH 4.0. The inhibition ability of CFS against the pathogens were not affected by the heating or protease treatment. However, pH modification in CFS to 6.5 resulted in an extreme reduction in their antifungal activity. These results may indicate that antifungal activities in CFS was caused by acidic compounds like PLA or organic acids rather than protein or peptide molecules.

• Applied Biological Chemistry
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• 제48권4호
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• pp.311-314
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• 2005

토양으로부터 분리된 약 1,200여주의 방선균으로부터 세포외로 phytase를 분비 생산하는 방선균 YB-26이 분리되었다. 분리균의 16S rRNA 염기서열을 조사한 결과 Streptomyces속에 속하는 균주의 서열과 상동성이 높았다. G.S.M 배지에서 분리균을 배양하여 얻은 배양 상등액을 ammonium sulfate 분획(15-70%), DEAE-Sepharose column 및 Q-Sepharose column 크로마토그래피를 하여 phytase를 부분 정제하였다. 부분정제된 phytase를 사용하여 효소반응을 실시한 결과 $60^{\circ}C$와 pH 7.0에서 최대활성을 보였으며, pH 6.0-8.0 범위에서 최대활성의 90%이상이 되는 활성을 나타냈다. 이 효소는 열안정성이 높지 않으며, $CaCl_2$의 존재하에서도 열안정성이 변화가 없는 것으로 확인되었다.

• Journal of Microbiology and Biotechnology
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• 제16권5호
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• pp.704-715
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• 2006

Oil spill was found in 1999 from a diesel storage facility located near the top of Baekun Mountain in Uiwang City. Application of bioremediation techniques was very relevant in removing oil spills in this site, because the geological condition was not amenable for other onsite remediation techniques. For efficient bioremediation, bacterial communities of the contaminated site and the uncontaminated control site were compared using both molecular and cultivation techniques. Soil bacterial populations were observed to be stimulated to grow in the soils contaminated with diesel hydrocarbon, whereas fungal and actinomycetes populations were decreased by diesel contamination. Most of the dieseldegrading bacteria isolated from contaminated forest soils were strains of Pseudomonas, Ralstonia, and Rhodococcus species. Denaturing gradient gel electrophoresis (DGGE) analysis revealed that the profiles were different among the three contaminated sites, whereas those of the control sites were identical to each other. Analysis of 16S rDNA sequences of dominant isolates and clones showed that the bacterial community was less diverse in the oil-contaminated site than at the control site. Sequence analysis of the alkane hydroxylase genes cloned from soil microbial DNAs indicated that their diversity and distribution were different between the contaminated site and the control site. The results indicated that diesel contamination exerted a strong selection on the indigenous microbial community in the contaminated site, leading to predominance of well-adapted microorganisms in concurrence with decrease of microbial diversity.

• Archives of Pharmacal Research
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• 제27권8호
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• pp.857-862
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• 2004

Naringenin, dietary flavonoid, is antioxidant constituents of many citrus fruits. In the present study, we investigated the effect of naringenin on 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-inducible CYP1 A 1 gene expression in mouse hepatoma Hepa-1c1c7 cells. Naringenin alone did not affect CYP1A1-specific 7-ethoxyresorufin O-deethylase (EROD) activity. In contrast, the TCDD-inducible EROD activities were markedly reduced upon concomitant treatment with TCDD and naringenin in a dose dependent manner. TCDD-induced CYP1A1 mRNA level was also markedly suppressed by naringenin. A transient transfection assay using dioxin-response element (DRE)-linked luciferase and electrophoretic mobility shift assay revealed that naringe-nin reduced transformation of the aryl hydrocarbons receptor(AhR) to a form capable of specif-ically binding to the DRE sequence in the promoter of the CYP1A1 gene. These results suggest the down regulation of the CYP1A1 gene expression by either naringenin in Hepa-1c1c7 cells might be antagonism of the DRE binding potential of nuclear AhR.