• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.84-89
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• 2005

Phosphomannomutase (PMM) is a key enzyme in prokaryotes and eukaryotes, which catalyzes the conversion of ${\alpha}$-D-mannose 6-phosphate to ${\alpha}$-D-mannose 1-phosphate. The latter is the substrate for the synthesis of GDP-mannose, which serves as the mannosyl donor for many metabolic pathways in the cells. We report here on the isolation of a gene from a genomic library of Sphingomonas chungbukensis DJ77, the pmmC gene encoding phosphomannomutase. The gene was cloned into E. coli expression vector, and the sequence was analyzed. The ribosomal binding site GGAAG lays 5 bp upstream of the ORF of 750 bp, which is initiated by ATG codon and terminated by TAG. The predicted sequence of the enzyme consists of 249 amino acids with a molecular mass of 27.4 kDa and showed $86.9\%$ similarity to that of eukaryotic phosphomannomutase after bioinformatical analyses with the conserved domain search of NCBI. The purified gene product revealed the activity of phosphomannomutase. In conclusion, we confirmed that pmmC gene encodes phosphomannomutase actually.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.458-465
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• 2021

Tubulysins are a group of bioactive secondary metabolites from myxobacteria exhibiting strong anticancer activity against various cancer cell lines. In this study, we describe the identification of putative tubulysin biosynthetic gene clusters (tubA~tubF) in the genome sequences of two tubulysin-producing myxobacterial strains, Archangium gephyra MEHO_002 and MEHO_004. The inactivation of the putative tubulysin biosynthetic genes resulted in a tubulysin-production defect. The DNA sequences of the A. gephyra MEHO_002 and MEHO_004 tubulysin biosynthetic genes were 97% identical, and the amino acid sequences of the encoded proteins shared a similarity of 97-100%. The nucleotide sequences of the tubulysin biosynthetic gene clusters in MEHO_002 and MEHO_004 were 86% identical to that in Cystobacter sp. SBCb004 known as a tubulysin-producing myxobacterium, and the organization of the clusters was identical except for the lack of a tubZ gene in the clusters in MEHO_002 and MEHO_004. The amino acid sequences of the proteins encoded by each gene were 88-97% similar to those encoded by SBCb004, and the domain compositions of the proteins were also identical.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.289-292
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• 2018

Using pyrene as the enrichment nutrient, a bacterial strain 10PY1A, was isolated by enrichment culture from oil-contaminated sea sand of Arabian Gulf in Saudi Arabia, and this strain belongs to the species Idiomarina piscisalsi, based on 16S RNA gene sequence analysis. The genome of I. piscisalsi strain 10PY1A contains 2,346 protein-coding sequences and an average GC content of 47.4% in its chromosome (2.59 Mbp). Genes encoding proteins related to the degradation of pyrene were existed in the strain 10PY1A genome, indicating that this strain can be used to degrade polycyclic aromatic hydrocarbons in oil-contaminated marine flora and soil.

• Microbiology and Biotechnology Letters
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• v.50 no.3
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• pp.395-403
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• 2022

In this study, we identified that the fermentation of Korean indigenous probiotics and red ginseng produced ginsenoside compound K (CK) from major ginsenosides. Based on whole genome sequencing of 19 probiotics species, β-glucosidase, α-arabinofuranosidase, β-xylosidase, and α-rhamnosidase related to bioconversion of ginsenosides are identified in the genome of 19 species, 3 species, 6 species, and 8 species, respectively. Among the 19 probiotics species, Bifidobacterium longum CBT BG7 converted from ginsenoside Rb1 to CK, and both B. breve CBT BR3 and B. lactis CBT BL3 converted ginsenoside Rb1 to Rd. The final concentration and yield of ginsenoside F2 and CK were higher in the fermentation with the nondisrupted cells than with disrupted cells. The combination of both CBT BG7 and BL3, and CBT BG7 and BR3 showed higher amounts of F2 than CBT BG7 only. CBT BG7 with adding α-amylase increased the amounts of F2. In this study, we identified that the fermentation of both Korean indigenous probiotic bacteria CBT BG7, BR3 and BL3, and red gingseng is able to produce CK, a bioactive compound that promotes health benefits.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.342-347
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• 2004

A bacterial strain YC4963 with antifungal activity against Colletotrichum orbiculare, a causal organism of cucumber anthracnose was isolated from the rhizosphere soil of Siegesbeckia pubescens Makino in Korea. Based on physiological and biochemical characteristics and 16S ribosomal DNA sequence analysis, the bac­terial strain was identified as Pseudomonas aurantiaca. The bacteria also inhibited mycelial growth of several plant fungal pathogens such as Botrytis cinerea, Fusarium oxysporum and Rhizoctonia solani on PDA and 0.1 TSA media. The antifungal activity was found from the culture filtrate of this isolate and the active compound was quantitatively bound to XAD adsorption resin. The antibiotic compound was purified and identified as phenazine-l-carboxylic acid on the basis of combined spectral and chemical analyses data. This is the first report on the production of phenazine-l-carboxylic acid by Pseudomonas aurantiaca.

• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.260-267
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• 2008

The choice of expression vector is very important for industrial production of proteins. Therefore, the systematic mining of promoters over a wider range of genetic resource and/or host is required. We previously reported a novel bidirectional reporting system (pBGR) for the isolation of promoters from metagenome and screened useful promoters that functioned constitutively in E. coli under general culture conditions. Among them, three promoter sequences including each upstream region were amplified by PCR and used to construct new expression vectors. To facilitate subcloning, a multi-cloning site was incorporated into the downstream region of the revere primer sequence. At these sites, GFP, esterase and $\beta$-glucosidase were subcloned and analyzed the constitutive expression ability of new promoter in terms of protein solubility and expression level. As a result, these vectors expressed the proteins constitutively to a level of $2{\sim}3%$ of the total cell protein in soluble fraction (>80 %). This study suggested that excavation of metagenomic promoters for construction of expression vector in a certain strain could provide a way for the development of the expression systems.

• Korean Journal of Microbiology
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• v.37 no.1
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• pp.28-36
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• 2001

These studies were carried out to understand the mechanisms of genes which are related in cell cycle progression at G1/S phase. Mutants involved in cell cycle progression and regulation in Saccharomyces cerevisiae were isolated and characterized. To isolate new mutants, we screened the sensitivity to ciclopirox olamine (CPO) which inhibits the cell cycle traverse at or very near the G1/S phase boundary in HeLa cell and budding yeast. As results, we isolated 30 mutants and named cos(ciclopirox olamine sensitivity: cos27∼cos57) mutants. To determine the phenotype of mutants, we examined the sensitivity to methyl-methane sulfonate (MMS) and hydroxyurea (HU). Several mutants were sensitive to MMS and HU. According to these Phenotypes, cos mutants were grouped into four. Group I mutants are cos27, cos28, cos32, cos33, cos36, cos37, cos40, cos42, cos46, cos50, cos52 and cos53 which show MMS, HU sensitivities and might act at a checkpoint pathway during S phase. Group II mutants are cos43 and cos48 which show MMS sensitivities and might act at a checkpoint pathway during Gl or G2 phase. Group III mutants are cos35, cos47, cos54, cos55 and cos56 which show HU sensitivities and might act at a progress pathway during S phase. Finally, Group IV mutants are cos29, cos30, cos31, cos34, cos38, cos39, cos41, cos44, cos45, cos49, cos51 and cos57 which show only CPO sensitivities. Moreover, we examined the terminal phenotype of mutants under fluorescent microscope and then found one of S phase checkpoint related mutant(cos37). Furthermore, we constructed the heterozygote strain between mutant and wild type haploid strains to study their genetic analysis of cos mutants.

• Korean Journal of Microbiology
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• v.47 no.1
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• pp.92-96
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• 2011

Rhodococcus sp. strain DK17 possesses three megaplasmids (380 kb pDK1, 330 kb pDK2, and 750 kb pDK3). The alkylbenzene-degrading genes (akbABCDEF) are present on pDK2 while the phthalate operons which are duplicated are present on both pDK2 (ophA'B'C'R') and pDK3 (ophABCR). DK17 with an optimal temperature of $30^{\circ}C$ showed no growth at $37^{\circ}C$. When transferred to $30^{\circ}C$, however, the $37^{\circ}C$ culture began to grow immediately, indicating that $37^{\circ}C$ is not lethal but stressful for DK17 growth. In addition, when exposed to $37^{\circ}C$ even for a short time, a part of DK17 cells lost the ability to degrade o-xylene (a model compound of alkylbenzenes). When two hundred colonies were randomly selected for colony PCR for pDK2-specific akbC, ophC', or pDK3-specific ophC, a total of 29 colonies were found to have lost at least one of the three genes. PFGE analysis clearly showed that all the mutants have different megaplasmid profiles from that of DK17 wild type, which are divided into five different cases: Type I (10 mutants, pDK2 loss and acquisition of a new ~700 kb plasmid), Type II (9 mutants, pDK2 loss), Type III (8 mutants, pDK3 loss and acquisition of a new ~400 kb plasmid), Type IV (1 mutant, pDK3 loss), and Type V (1 mutant, pDK2 and pDK3 loss and acquisition of the ~400 kb and ~700 kb plasmids). The above results showing that growth temperature changes can induce physical changes in bacterial genomes suggest that environmental changes in habitats including temperature fluctuations affect significantly the evolution of bacteria.

• Microbiology and Biotechnology Letters
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• v.51 no.1
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• pp.83-92
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• 2023

In this study, bioconversion of rutin to quercetin was confirmed by the fermentation of Korean indigenous probiotics and tartary buckwheat. Based on whole genome sequencing of 17 probiotics species, α-rhamnosidase, related to bioconversion of isoquercetin (quercetin 3-β-D glucoside) from rutin, is identified in the genome of CBT BG7, LC5, LR5, LP3, LA1, and LGA1. β-Glucosidase, related to bioconversion of isoquercetin to quercetin, is identified in the genome of all 17 species. Among the 17 probiotics species, 6 probiotics including CBT BG7, LR5, LP3, LA1, LGA1 and ST3 performed the bioconversion of rutin to quercetin up to 21.5 ± 0.3% at 7 days after fermentation. The fermentation of each probiotics together with enzyme complex Cellulase KN^® was conducted to reduce the time of bioconversion. As a result, CBT LA1 which showed the highest yield of bioconversion of 21.5 ± 0.3% when the enzyme complex was not added showed high bioconversion yield of 84.6 ± 0.5% with adding the enzyme complex at 1 day after fermentation. In particular, CBT ST3 (96.2 ± 0.4%), SL6 (90.1 ± 1.4%) and LP3 (90.0 ± 0.4%) showed high yield of bioconversion more than 90%. In addition, such probiotics including high levels in quercetin indicated the inhibitory effects of NO production in LPS-induced RAW264.7 cells. In this study, we confirmed that the fermentation of Korean indigenous probiotics and enzyme complex together with roasted tartary buckwheat increased the content of quercetin and reduced the time of bioconversion of rutin to quercetin which is a bioactive compound related to anti-inflammatory, antioxidants, anti-obesity, and anti-diabetes.

• Proceedings of the KSMRM Conference
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• 2002.11a
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• pp.50-56
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• 2002

최근 3-4년간 MR의 hardware와 software의 급격한 발전으로 마침내 작년 말에 3.0 Tesla whole body MR 장비가 미국 FDA의 공인을 받았다. 한국에서도 일찍부터 3T MR장비의 개발이 이루어 졌고 이미 설치되어 연구와 임상이 이용되고 있다. 여러 회사에서 개발 및 연구된 전신 3.0T MR 장비가 여러가지 가능성을 보이고 임상 도입 단계에 있지만 아직까지 실지 임상에서는 뇌신경계 분야가 주류를 이루고 있다. 지금 뇌신경계 분야에서 보편적으로 늘리 사용되고 있는 1.5T MR 장비는 모든 면에서 상당히 안정적으로 임상 및 연구에 이용되고 있다. 1-2년 전만 해도 3.0 T MR기기는 뇌신경계 영역에서도 임상적으로 늘리 사용되기에는 안정적인 면에서는 1.5T 기기에 비해서 떨어지는 것이 사실이었다. 그래서 주로 연구실 영역에서 많이 이용되고 있었다. 그러나 지금 본원에 설치 완료되어 임상에 적용한지 6개월 정도 이용한 예에서 보면 (about 2300 cases/6months) hardware, software적인 면에서 아직 조금의 불편함이 있지만 많은 부분이 충분히 인지되고 개선이 가능한 부분으로 거의 불편함이 사라질 것으로 기대되고 있고, 불편함을 넘을 수 있는 여러 가지 장점이 있다고 본다. 고자장 (>3.0 T) MRI의 매력은 자장에 비례적으로 SNR, spectral resolution이 높아지고, T1, BOLD등에 의한 대조도가 향상한다는 것이다. SNR의 증가는 temporal, spatial 분해능을 증가시키고, spectral resolution이 높아짐에 따라 MR spectroscopy상에서 주요 대사물질 이외 작은 대사물질에 관한 스펙트럼의 분석을 향상시킨다. 이처럼 고자장 MR은 근본적인 장점을 가지고 있고 이러한 장점이 고자장 MR 시대로 가야 할 이유을 모두 설명하고 있다고 생각된다.세포질등이 있으며, 이들중에서 lysosomes, peroxisomes, 그리고 미토콘드리아가 특정한 유전성 백질질환에 중요한 역할을 하는 것이 밝혀졌다. 이러한 질환들은 최소한 각 소기관에 의한 질환군으로 분류될 수 있다.SXR이 ER의 transactivation 효과를 약간 촉진한 반면 MDA-MB-231세포는 SXR을 제외한 CAR와 PPAR${\gamma}$에 의해 ER의 transactivation 효과가 약간 증가되는 경향을 보였다. 이러한 결과는 유방암세포에서는 CAR, SXR, PPAR${\gamma}$과 같은 xenobiotic nuclear receptor에 의한 ER transactivation 효과가 간암세포와는 다르게 나타나며, 유방암의 종류에 따라서 endogenous CAR, SXR, PPAR${\gamma}$수용체가 다르게 발현됨으로써 이들에 대한 반응이 서로 상이한 특징을 나타낼 수 있을 것으로 사료된다. 따라서 estrogen receptor에 의해 매개되는 estrogn의 전사활성조절기전이 표적세포에 따라 다른 경로를 포함 할 수 있음을 시사한다.서 흡착 능력이 우수하게 나타났으며, 황화수소는 펄라이트, 왕겨, 소나무수피에서 상대적으로 우수한 것으로 나타났으며, 혼합충전재는 암모니아의 경우 코코넛과 펄라이트의 비율이 7：3인 혼합 재료 3번과 소나무수피와 펄라이트의 비율이 7：3인 혼합 재료 6번에서 다른 혼합 재료에 비하여 우수한 것으로 나타났다. 4. 코코넛과 소나무수피의 경우 암모니아 가스에 대한 흡착 능력은 거의 비슷한 것으로 사료되며, 코코넛의 경우 전량을 수입에 의존하고 있다는 점에서 국내 조달이 용이하며, 구입 비용도 적게 소요되는 소나무수피를 사용하는 것이 경제적이라고 사료된다. 5. 마지막으로 악취제거 미생물균주를 접종한 소나무수피 50%와 펄라이트