• Journal of Microbiology and Biotechnology
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• 제26권4호
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• pp.666-674
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• 2016

The bacterial strains were screened as potential starters for fermenting low-salt doenjang (a Korean traditional fermented soybean paste) using Korean doenjang based on proteolytic and antipathogenic activities under 6.5-7.5% NaCl conditions. Phylogenetic analysis based on 16S rRNA gene sequences showed that they all belonged to the genus Bacillus. Proteolytic and antipathogenic activities against Escherichia coli, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, and Aspergillus flavus, as well as fibrinolytic, amylase, and cellulase activities of the 10 strains were quantitatively evaluated. Of these, strains D2-2, JJ-D34, and D12-5 were selected, based on their activities. The functional, phenotypic, and safety-related characteristics of these three strains were additionally investigated and strains D2-2 and D12-5, which lacked antibiotic resistance, were finally selected. Strains D2-2 and D12-5 produced poly-γ-glutamic acid and showed various enzyme activities, including α-glucosidase and β-glucosidase. Growth properties of strains D2-2 and D12-5 included wide temperature and pH ranges, growth in up to 16% NaCl, and weak anaerobic growth, suggesting that they facilitate low-salt doenjang fermentation. Strains D2-2 and D12-5 were not hemolytic, carried no toxin genes, and did not produce biogenic amines. These results suggest that strains D2-2 and D12-5 can serve as appropriate starter cultures for fermenting low-salt doenjang with high quality and safety.

• 대한임상검사과학회지
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• 제48권3호
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• pp.217-224
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• 2016

Acinetobacter baumannii는 병원환경에 광범위하게 분포하고 있으며, 원내감염의 중요한 원인균으로 병원에서 집단감염 일으키고, 중증의 기저질환을 가진 환자에게 감염되면 감염환자의 사망률이 다른 환자에 비해 월등히 높은 것으로 알려져 있다. 본 연구는 충청남도 천안시에 소재한 대학병원 두 곳의 진단검사의학과에 의뢰된 가검물에서 분리한 다제 내성 A. baumannii 85주의 항생제 내성률과 내성유전자 양상에 대해 조사하였다. Carbapenemase와 Class B ${\beta}$-lactamase의 생성균주를 선별하기 위하여 modified Hodge test (MHT)와 IMP-EDTA double-disk synergy test를 실시하였다. 항생제 내성을 유발하는 carbapenemases, 16S rRNA methylases, aminoglycoside-modifying enzymes (AMEs)을 확인하기 위하여 PCR을 시행 하였으며, A. baumannii가 분리된 두 대학병원간 분리균주의 유전학적인 근연도를 확인하기 위해 REP-PCR을 시행하였다. 실험결과 3균주를 제외한 82균주(96.5%)에서 $bla_{OXA-23-like}$과 $bla_{OXA-51-like}$이 검출되었다. $bla_{OXA-23-like}$ 유전자가 검출된 균주에서는 $bla_{OXA-23-like}$ 유전자 상부에 ISAba1 유전자가 확인되어 carbapenemase에 대한 내성을 유도하는 것으로 확인 되었다. Aminoglycoside에 대한 내성을 유발하는 16S rRNA methylase 유전자인 armA는 A병원에서 분리한 38균주 중 34균주(89.5%)에서, B병원에서 분리한 47균주 중 40균주(85.1%)에서 확인되었고, aminoglycoside modifying emzyme 유전자는 A병원 유래 38 균주 중 33 균주(70.2%)에서, B병원 유래 47균주 중 44 균주(93.6%)에서 aac(3)-IIa/ant(2")-Ia/aac(6')-Ib가 확인됨에 따라 천안지역에서 분리되는 대부분의 다제 내성 A. baumannii 균주는 acethyltransferase와 adenyltransferase를 동시에 발현하는 것을 알 수 있었다. 본 실험 결과는 충청남도 천안시에서 분리된 MDR A. baumannii를 대상으로 한 보고서로서, 항생제 내성유형과 내성 유전자의 분포를 확인하여 MDR A. baumannii 균주에 의한 감염증의 치료 지침과 내성세균 확산 방지를 위해 필요한 기초 자료로 활용할 수 있을 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제30권9호
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• pp.1367-1378
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• 2020

The polyphagous eri silk moth, Samia ricini, is associated with various symbiotic gut bacteria believed to provide several benefits to the host. The larvae of S. ricini were subjected to isolation of gut bacteria using culture-dependent 16S rRNA generic characterization, metagenomics analysis and qualitative enzymatic assays. Sixty culturable aerobic gut bacterial isolates comprising Firmicutes (54%) and Proteobacteria (46%); and twelve culturable facultative anaerobic bacteria comprising Proteobacteria (92%) and Firmicutes (8%) were identified inhabiting the gut of S. ricini. The results of metagenomics analysis revealed the presence of a diverse community of both culturable and un-culturable gut bacteria belonging to Proteobacteria (60%) and Firmicutes (20%) associated with seven orders. An analysis of the results of culturable isolation indicates that these bacterial isolates inhabited all the three compartments of the gut. Investigation on persistence of bacteria coupled with metagenomics analysis of the fifth instar suggested that bacteria persist in the gut across the different instar stages. In addition, enzymatic assays indicated that 48 and 75% of culturable aerobic, and 75% of anaerobic gut bacterial isolates had cellulolytic, lipolytic and nitrate reductase activities, thus suggesting that they may be involved in food digestion and nutritional provision to the host. These bacterial isolates may be good sources for profiling novel genes and biomolecules for biotechnological application.

• 한국축산식품학회지
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• 제36권6호
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• pp.779-786
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• 2016

This study aimed to evaluate the prevalence of Listeria monocytogenes on livestock farms in Korea and determine their serotypes and genetic correlations. Twenty-five livestock farms in Korea (central: 15, south west: 7, south east: 3) were visited 2-3 times, and 2,018 samples (feces: 677, soil: 680, silage: 647, sludge: 14) were collected. Samples were enriched in LEB (Listeria enrichment broth) and Fraser broth media, and then plated on Palcam agar. The isolates were identified by PCR and 16S rRNA gene sequencing. Then, the sero-types, presence of virulence genes (actA, inlA, inlB, plcB, and hlyA), and antibiotic resistance were determined. Genetic correlations among the isolates were evaluated by analyzing the restriction digest pattern with AscI. Of the 2,018 samples, only 3 (0.15%) soil samples (FI-1-FI-3) from 1 farm in the south east region were positive for L. monocytogenes. Based on biochemical tests and multiplex PCR, the serotype of the isolates were 4ab (FI-1 and FI-3) and 3a (FI-2), which are not common in foodborne L. monocytogenes. The 3a sero-type isolate was positive for all tested virulence genes, whereas the 4ab serotype isolates were only positive for hlyA, actA, and inlA. The isolates were resistant to all 12 tested antibiotics, especially FI-3. The genetic correlations among the isolates were 100% for those of the same serotype and 26.3% for those of different serotypes. These results indicate that the prevalence of L. monocytogenes on livestock farms in Korea is low; however, the isolates are pathogenic and antibiotic resistant.

• 미생물학회지
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• 제54권3호
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• pp.289-292
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• 2018

석유로 오염된 사우디아라비아의 바다 모래에 pyrene을 영양물로 첨가하는 집식배양을 통해 우점하는 박테리아 균주 10PY1A가 분리되었다. 이 균주는 16S rRNA 유전자 분석을 통해 Idiomarina piscisalsi로 동정되었다. 이 균주는 G + C 비율이 47.4%이며 2.69 Mbp 크기의 원형 염색체를 보유하고 있으며, 해당 염색체는 다환방향족탄화수소 분해와 관련 있는 유전자를 포함한 2,346개의 단백질 코딩 유전자로 구성되어 있다. 이는 이 균주가 석유로 오염된 해양과 토양에서 방향족탄화수소를 분해하는 데 사용될 수 있음을 보여준다.

• Journal of Microbiology and Biotechnology
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• 제27권10호
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• pp.1744-1752
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• 2017

Myeolchi jeotgals (MJs) were prepared with purified salt (PS), solar salt aged for 1 year (SS), and bamboo salt (BS) melted 3 times at 10% and 20% (w/w) concentrations, and fermented for 28 weeks at $15^{\circ}C$. BS MJ showed higher pH and lower titratable acidities than the other samples because of the alkalinity of bamboo salt. Lactic acid bacteria counts increased until 4-6 weeks and then decreased gradually, and were not detected after 20 weeks from MJs with 10% salt. Yeast counts of PS MJs were higher than those of BS and SS MJs. Bacilli were detected in relatively higher numbers throughout the 28 weeks, like marine bacteria, but archae were detected in lower numbers during the first 10 weeks. When 16S rRNA genes were amplified from total DNA from PS MJ (10% salt) at 12 weeks, Tetragenococcus halophilus was the major species. However, Staphylococcus epidermidis was the dominant species for BS MJ at the same time point. In SS MJ, T. halophilus was the dominant species and S. epidermidis was the next dominant species. BS and SS MJs showed higher amino-type nitrogen, ammonia-type nitrogen, and volatile basic nitrogen contents than PS MJs. SS and BS were better than PS for the production of high-quality MJs.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.178-184
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• 2015

This work aims to utilize wastes from the potato starch industry to produce single-cell protein (SCP) with high lysine content as animal feed. In this work, S-(2-aminoethyl)-_L-cysteine hydrochloride-resistant Bacillus pumilus E1 was used to produce SCP with high lysine content, whereas Aspergillus niger was used to degrade cellulose biomass and Candida utilis was used to improve the smell and palatability of the feed. An orthogonal design was used to optimize the process of fermentation for maximal lysine content. The optimum fermentation conditions were as follows: temperature of 40℃, substrate concentration of 3%, and natural pH of about 7.0. For unsterilized potato starch wastes, the microbial communities in the fermentation process were determined by terminal restriction fragment length polymorphism analysis of bacterial 16S rRNA genes. Results showed that the dominant population was Bacillus sp. The protein quality as well as the amino acid profile of the final product was found to be significantly higher compared with the untreated waste product at day 0. Additionally, acute toxicity test showed that the SCP product was non-toxic, indicating that it can be used for commercial processing.

• 식물병연구
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• 제28권3호
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• pp.172-178
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• 2022

The graft cactus (Gymnocalycium mihanovichii) continues to be exported to more than 20 countries worldwide. In April 2021, typical bacterial symptoms of soft rot were observed in the graft cactus (cv. Yeonbit) in Goyang, Gyeonggi-do, Korea, resulting in economic losses in cactus production. The stems turned dark brown and the flowers were covered with black rot. The bacterial strain designated as KNUB-01-21 was isolated from infected stems and flowers. The results of the morphological and biochemical tests of the isolate were similar to those of Pectobacterium brasiliense. For molecular analysis, the 16S rRNA region and three housekeeping genes (dnaX, leuS, and recA) of the strain KNUB-01-21 were amplified. Based on the results of the molecular analysis and morphological and biochemical tests, KNUB-01-21 was identified as P. brasiliense. The pathogenicity of KNUB-01-21 on graft cactus was confirmed by an inoculation test. Artificial inoculation using P. brasiliense KNUB-01-21 produced soft rot symptoms on the grafted cactus, and the same bacterium was re-isolated and re-identified. This is the first report of P. brasiliense causing soft rot in graft cactus in Korea.

• 한국임상수의학회지
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• 제32권2호
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• pp.158-161
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• 2015

세균성 피부염은 항생제 치료가 필요한 흔한 질환이다. 최근 항생제 내성 세균이 전 세계적으로 증가하고 있는 추세이다. 본 연구에서는 개 피부염 병변에서 분리한 S. pseudintermedius의 항생제 내성 유전자 발현을 조사하고, 이를 디스크 감수성 시험결과와 비교하였다. 총 7개의 S. pseudintermedius 균주가 연구에 포함되었다. 세균 동정은 16S rDNA 염기 서열 분석으로 실시하였다. 조사 결과 분리한 S. pseudintermedius 모두가 1개 이상의 항생제 내성유전자(mecA, blaZ and aac[6']/aph[2"])를 보유하였다. 전체 분리균주가 blaZ 유전자에 양성을 보였으나, 두 개 균주만이 amoxicillin/clavulanate에 내성을 나타냈다. Gentamicin 내성을 나타낸 5개 균주 중, 1개는 aac(6')/aph(2") 유전자에 대한 PCR 검사에서 음성을 나타냈다. 이 결과는 내성 유전자 검출 PCR 기법과 디스크 감수성 시험 기법이 항생제 내성을 검사하는 데에 있어 상호보완적인 관계를 가짐을 제시한다.

• 미생물학회지
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• 제49권1호
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• pp.46-50
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• 2013

일부 근권세균은 ACC deaminase를 생성하여 식물의 생장을 저해하고 노화를 촉진시키는 식물호르몬 에틸렌의 수준을 낮춤으로써 스트레스 조건 하의 식물의 생장을 지속시킨다. 본 연구에서는 모래사장에서 자라는 식물의 근권에서 ACC deaminase를 생성하는 세균 균주들을 분리하여 16S rDNA 염기서열 분석을 통해 Escherichia hermannii m-2, Enterobacter asburiaem-4, Pseudomonas thivervalensis BD2-26, and Pseudomonas brassicacearum subsp. neoaurantiaca BD3-35로 동정하였다. BD3-35 균주는 이들 중 가장 높은 ACC deaminase 활성, 20.26 ${\alpha}$-ketobutyrate ${\mu}M/mg$ protein/h을 나타내었다. 균주 BD3-35와 BD2-26는 식물호르몬 시토키닌, m-4는 옥신 IAA와 IBA, 그리고 균주 m-2는 ABA 생성능을 가졌다. 이 균주들은 모두 토마토종자 발아 시 유묘의 뿌리신장을 유의성 있게 촉진하였다. 또한 7일간 자란 토마토 식물에 처리하고 가뭄 스트레스 하에서 7일간 재배하였을 때 비접종 대조군에 비해 균주 BD3-35, m-2와 m-4는 토마토 뿌리의 길이를 각각 14, 15와 35% 증가시켰으며, m-2, BD2-26와 BD3-35는 토마토 식물의 건조중량을 각각 22, 33과 68% 증가시켰다. 따라서 이 균주들은 가뭄 스트레스 하의식물을 위한 미생물 비료로 사용될 수 있는 가능성을 보였다.