• Applied Biological Chemistry
• /
• 제30권1호
• /
• pp.40-45
• /
• 1987

Alcaligenes aquamarinus에 의한 각종 PCBs와 유기염소계 농약 및 토양중의 PBC-42 분해 등을 검토하고 TLC 등으로 분해산물을 검정한 결과 PCBs에서는 염소화도가 낮은 PCBs가 더 쉽게 분해되었고 유기염소계 농약에서는 p,p'-DDT>r-BHC>Thiolix 순으로 분해되었다. 토양중의 PCB-42의 분해는 살균하지 않은 토양시료에서 $25^{\circ}C$, 25일 배양으로 약 40%가 분해되었다. PCB-42의 분해로 생성된 황색물질의 최대 흡수파장은 pH 4.5에서 258nm와 295nm이었고 분해산물의 일종인 p-chlorobenzoic acid가 TLC와 GC에 의하여 확인되었다.

• Mass Spectrometry Letters
• /
• 제9권3호
• /
• pp.81-85
• /
• 2018

In this study, the chemical composition of bacterial melanin isolated from the Streptomyces glaucescens strain was elucidated by ultra-high-resolution Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry. Ultra-high-resolution mass profiles of the microbial melanin product were acquired using a 15 Tesla FT-ICR mass spectrometer in positive and negative ion modes via electrospray ionization to obtain more complete descriptions of the molecular compositions of melanin-derived organic constituents. A mass resolving power of 500,000 (at m/z 400) was achieved for all spectra while collecting 400 scans per sample with a 4 M transient. The results of this analysis revealed that the melanin pigment isolated from S. glaucescens predominantly exhibits CHON and CHO species, which belong to the proteins class of compounds, with the mean C/O and C/N ratios of 4.3 and 13.1, thus suggesting that the melanin could be eumelanin. This analytical approach could be utilized to investigate the molecular compositions of a variety of natural or synthetic melanins. The compositional features of melanins are important for understanding their formation mechanisms and physico-chemical properties.

• Journal of Microbiology and Biotechnology
• /
• 제12권2호
• /
• pp.286-291
• /
• 2002

Histone deacetylase I (HDAC1) works as one of the components in a nucleosome remodeling (NuRD) complex that consists of several proteins, including metastasis-associated protein 1 (MTA1). Since the protein-protein interaction of HDAC1 and MTA1 would appear to be important for both the integrity and functionality of the HDAC1 complex, the interruption of the HDAC1 and MTA1 interaction may be an efficient way to regulate the biological function of the HDAC1 complex. Based on this idea, a yeast two-hybrid system was constructed with HDAC1 and MTA1 expressing vectors in the DNA binding and activation domains, respectively. To verify the efficiency of the assay system, 3,500 microbial metabolite libraries were tested using the paper disc method, and KB0699 was found to inhibit the HDAC1 and MTA1 interaction without any toxicity to the wild-type yeast. Furthermore, KB0699 blocked the interaction of HDAC1 and MTA1 in an in vitro GST pull down assay and induced morphological changes in B16/BL6 melanoma cells, indicating the interruption of the HDAC1 complex function. Accordingly, these results demonstrated that the yeast assay strain developed in this study could be a valuable tool for the isolation of a HDAC1 complex disruptor.

• 한국미생물·생명공학회지
• /
• 제23권6호
• /
• pp.641-646
• /
• 1995

During the screening of inhibitors of melanin biosynthesis from microbial secondary metabolites, a fungal strain MR304 which was capable of producing high level of an inhibitor was selected. Based on taxonomic studies, this fungus could be classified as Trichoderma harzianum. The active compound (MR304-1) was purified from culture broth by Diaion HP-20 column chromatography, ethylacetate extraction, Sephadex LH-20 column chromatographv and HPLC. The inhibitor was identified as 3-(1,5-dihvdroxy-3-isocyanocyclopent-(E)-3-envl)prop-2-enoate by spectroscopic methods of UV, ESIMS, $^{1}$H-NMR, $^{13}$C-NMR, NOE, HMQC and HMBC. MR304-1 showed strong mushroom tyrosinase inhibitory activity with IC$_{50}$ value of 0.25 $\mu $g/ml. It inhibited melanin biosynthesis with 15 mm inhibition zone at 30 $\mu $g/paper disc in Streptomyces bikiniensis, a bacterium used as an indicator organism in this work. It also inhibited melanin biosynthesis in B16 melanoma cells with a niinimum inhibitory concentration of 0.05 $\mu $g/ml.

• Journal of Microbiology and Biotechnology
• /
• 제25권6호
• /
• pp.887-892
• /
• 2015

Uricase is an important microbial enzyme that can be used in the clinical treatment of gout, hyperuricemia, and tumor lysis syndrome. A total of 127 clinical isolates of Pseudomonas aeruginosa were tested for uricase production. A Pseudomonas strain named Ps43 showed the highest level of native uricase enzyme expression. The open reading frame of the uricase enzyme was amplified from Ps43 and cloned into the expression vector pRSET-B. Uricase was expressed using E. coli BL21 (DE3). The ORF was sequenced and assigned GenBank Accession No. KJ718888. The nucleotide sequence analysis was identical to the coding sequence of uricase gene puuDof P. aeruginosa PAO1. We report the successful expression of P. aeruginosa uricase in Escherichia coli. E. coli showed an induced protein with a molecular mass of about 58 kDa that was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. We also established efficient protein purification using the Ni-Sepharose column with activity of the purified enzyme of 2.16 IU and a 2-fold increase in the specific activity of the pure enzyme compared with the crude enzyme.

• Journal of Microbiology and Biotechnology
• /
• 제19권9호
• /
• pp.881-887
• /
• 2009

Forty-four eicosapentaenoic acid (EPA)-producing microbial strains were isolated from the intestines of marine fishes. Among them, one strain showing a maximum level of EPA (4.78% of total fatty acids) was identified as Shewanella sp. BR-2 on the basis of its 168 rRNA sequence. The EPA content reached a maximum level during the mid-exponential phase of cell growth, and gradually decreased with further growth of the cells. A cosmid DNA including the EPA biosynthesis gene cluster consisting of pfaA-E was isolated from a cosmid library of genomic DNA of Shewanella sp. BR-2, named pCosEPA-BR2. An E. coli clone harboring pCosEPA-BR2 produced EPA at a maximum level of 7.5% of total fatty acids, confirming the EPA biosynthesis activity of the cloned gene cluster.

• Journal of Microbiology and Biotechnology
• /
• 제30권7호
• /
• pp.1082-1091
• /
• 2020

Microbial transglutaminases (MTGs) are widely used in the food industry. In this study, the MTG gene of Streptomyces sp. TYQ1024 was cloned and expressed in a food-grade bacterial strain, Bacillus subtilis SCK6. Extracellular activity of the MTG after codon and signal peptide (SP Ync M) optimization was 20 times that of the pre-optimized enzyme. After purification, the molecular weight of the MTG was 38 kDa and the specific activity was 63.75 U/mg. The optimal temperature and pH for the recombinant MTG activity were 50℃ and 8.0, respectively. MTG activity increased 1.42-fold in the presence of β-ME and 1.6-fold in the presence of DTT. Moreover, 18% sodium chloride still resulted in 83% enzyme activity, which showed good salt tolerance. Cross-linking gelatin with the MTG increased the strength of gelatin 1.67 times and increased the thermal denaturation temperature from 61.8 to 75.8℃. The MTG also significantly increased the strength and thermal stability of gelatin. These characteristics demonstrated the huge commercial potential of MTG, such as for applications in salted protein foods.

• 미생물학회지
• /
• 제15권4호
• /
• pp.159-169
• /
• 1977

The bacteria of red colonies isolated from soil were identified as Serratia marcescens. The best solvent for pigment extraction was n-buthanol and the pigment was identified as prodigiosene. The extracted pigment was stable on temperature and light but not on acidity. The redpigment color changed into red in alkaline solution. The maximum absorbancy of pigment was 466 nm in alkaline condition and 540 nm in acid condition. And the pigment formed single spot on the TLC(starch). By the result of infra red spectrum, the red pigment has the same absorption pattern comparing with, the prodigisin produced by S. marcescens strain Nima. It was confirmed that the pigment was secondary metabolite and that the maximal peak of production appeared at 30 hrs after the inoculation, when the bacterial growth was in statinary state. Referring to the effect of temperature, the pigment was not formed at $36^{\circ}C$ and the optimal temperature for both of bactrial growth and pigmentation was $30^{\circ}C$. The optimal range of pH for pigmentation was 5.0 and under the condition the bacterial growth was not affected at all. Examining the effects of light, the bacterial pigment ation was more increased in darkness than in visible light.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
• /
• pp.540-543
• /
• 2000

본연구에서는 Rhodococcus rhodochrous IGTS8 균주를 사용하여 고농도의 유상에서 탈황효율을 높이기 위한 오일 함유비, pH, 영양물질의 영향을 조사하였다. 오일 함유비에 의한 영향은 유상이 30%이하일 경우 그리 크지 않았으며, pH 조절에 의해 50%, 영양물질의 강화에 의해 32%의 탈황효율이 증가했다. 강화배지에서 pH를 조절하며 배양한 결과, 기존의 배양에 비해 136% 탈황효율이 증가했다.

• KSBB Journal
• /
• 제11권4호
• /
• pp.470-475
• /
• 1996

썩은 나무 잎에서 세포외 생물고분자 응집제를 생 산하는 흰 곰팡이를 분리하여 Pestalotiopsis sp. M01로 동정하였다. 이 균주가 생산하는 생물고분자 응집제의 응집활성에 대한 배양 조건을 무기염 배지인 Czapek-Dox 배지를 기초로 하여 탄소원, 질소 원, 배지의 pH멸, 그리고 옹도별로 조사하였다. 그 결과로서 응집활성 조건은 3 % sucrose, 0.3 % $KN0_3$, pH 7, 그리고 $25^{\circ}C$에서 최대 응집활성을 나타내었다. 반면에 이 균주의 생육 조건은 3% su-crose, 0.3 % $KN0_3$, pH 5, 그리고 $25^{\circ}C$에서 최대 생육을 나타내었다.