• Applied Biological Chemistry
• /
• 제39권2호
• /
• pp.99-103
• /
• 1996

토양으로부터 inulin을 기질로 하여 중합도가 큰 oligo당으로 가수분해하는 새로운 endo형 Inulinase 생산균주로, Arthrobacter sp. S37를 분리, 선발하였다. 본 효소의 생산은 inulin과 돼지감자 추출물에 의해 유도되었으며, 탄소원으로 1.5% 돼지감자 추출물, 유기질소원으로 1.0% yeast extract, 무기 질소원으로 0.5% $NaNO_3$를 사용하였을 때 효소 생산이 가장 많았다. 또한 최적 배양 pH와 온도는 각각 pH 8.0과 $30^{\circ}C$로 나타났다. 최적 배지와 최적 배양 조건하에서 배양한 결과 배양 24시간에 10.8 units/ml로 최대효소생산을 보였다.

• 한국해양바이오학회지
• /
• 제1권4호
• /
• pp.275-281
• /
• 2006

3.4-dichloroaniline (DCA)를 함유한 최소배지에서의 집식배양과 배양 후 HPLC에 의한 잔류분석을 통해 3,4-DCA의 분해 능력이 우수한 균주 Arthrobacter sp. YM-14를 여천석유화학공단 인근의 갯벌에서 분리하였다. 분리균 YM-14는 1/10 LB 배지에 함유된 50 ppm의 3,4-DCA를 12 시간 만에 완전히 제거하였다. 이외에도 분리균 YM-14는 3-chloroaniline(CA), 2,5-DCA 및 3,5-DCA의 분해 활성을 나타내었으나 2-CA, 4-CA와 2,4-DCA에 대한 분해활성을 가지고 있지는 않았다. 또한, 분리균 YM-14에서 3,4-DCA의 유도에 의한 catechol 1,2-dioxygenase 활성의 증가가 관찰되었다. 이러한 결과는 catechol 1,2-dioxygenase이 3,4-DCA 분해에 관여하는 중요한 효소군중의 하나로 생각된다.

• 미생물학회지
• /
• 제23권3호
• /
• pp.177-183
• /
• 1985

20-80%로 황산암모늄 분획한 효소액을 사용하여 Arthrobacter sp. JH-13균주가 생산하는 세포외 cytosme deaminase의 성질을 검토하였다. 검토한 기질 중, 본 효소는 cytosine과 5- f1uorocytosine을 기질로 이용하였으며, 효소의 활성에 대한 최적 pH는 8.0부근이었고, 최적 온도는 $40^{\circ}C$부근으로 나타났다. 본 효소는 0.2M의 p potassium phosphate 완충액 (pH 8.0) 보다 0.2M의 tris-HCl 완충액 (pH 8. 0) 에서 더욱 안정하였다. 온도에 대한 안정성을 검토한 결과. $50^{\circ}C$ 부근까지는 대체로 안정하였으나, $70^{\circ}C$에서는 완전히 활성을 잃었다. 또한 ImM의 $Fe^{3+},\;K^+\;Na^+$ 이온은 효소의 활성을 증가시켰으나 0.01mM의 $Co^{2+},\;Cu^{2+},\;Ni^{2+},\;Hg^{2+},\;Ag^{2+},\;Zn^{2+},\;Ba^{2+},\;Mg^{2+}$ 이온들은 효소의 활성올 강력하게 저해하였다. O.lmM의 p-mercuribenzoate, trichloroacetic acid, N-ethylmaleim mide등은 효소의 활성을 완전히 저해하였으며. O.lmM의 2-mercaptoethanol은 효소의 활성올 약간 증가시키는 것으로 나타났다.

• 한국환경과학회지
• /
• 제15권10호
• /
• pp.983-988
• /
• 2006

The textile remains have been affected largely by environmental factors including microorganisms because they were composed of organic compounds to be easy to damage. So, we selected 8 strains of the 131 isolated strains from museum environments and textile remains by high pretense activity, and identified them for measuring the antibacterial activity of Gingko biloba extracts. They were identified Genus Arthrobacter spp. 3 strains (Arthrobacter nicotiannae A12, Arthrobacter sp B12, Arthrobacter oxidans B13), Cenus Bacillus spp. 2 strains (Bacillus licheniformis D9, Bacillus cereus D33), Genus Pseudomonas spp. 2 strains (Pseudomonas putida A24, Pseufomonas fluorescene C21) and a Genus Staphylococcus sp. 1 strain (Staphylococcus pasteuri D3) as closest strains through the blast search of NCBI. Though antibacterial activity of the extracts of Gingko biloba leaves as MIC was lower than that of other pharmaceutical antibiotics. However the extracts was crude extracts, the extracts might have good antibacterial against most of the isolates from museum. Especially, the antifungal activity of Gingko biloba is known previously, the extracts of Gingko biloba leaves has possibility of usage as a good natural material for conservation of remains.

• Journal of Microbiology and Biotechnology
• /
• 제4권2호
• /
• pp.119-125
• /
• 1994

Steroid $\Delta^1$-dehydrogenase purified from hydrocortisone-induced cells of Arthrobacter simplex converted various 3-ketosteroids into their corresponding $\Delta^1$-dehydrogenated products. The transformation efficiencies depend upon the chemical structure of the steroids, especially length of the side chain at 17 position and hydroxyl groups at 11 and 17 positions. The Km values for androstenedione, the most favorable substrate examined, and hydrocortisone were 74 ${\mu}M$ and 294 ${\mu}M$, respectively. The optimum temperature and pH of the enzyme reaction were 35$^{\circ}C$ and pH 9, respectively, and the enzyme was relatively stable at the range from 20 to 35$^{\circ}C$ and from pH 5 to 10 after one hour of incubation. The enzyme activity was markedly inhibited in the presence of $Cu^{2+},\;Fe^{3+},\;Hg^{2+},\;Mo^{6+}$ ions, and somewhat inhibited by $Zn^{2+}$ and $Fe^{2+}$. $\alpha,\alpha'$-Dipyridyl that inhibits 9$\alpha$-hydroxylase and accumulates 1,4-androstadiene-3,17-dione from sterols revealed no inhibitory effect on this enzyme. EGTA showed inhibitory effect. $\beta$-Estradiol competitively inhibited the enzyme activity. Chemical modifications of the enzyme were attempted with several reagents. p-Hydroxymer-curibenzoate showed inhibition of the enzyme activity and protection of the substrate. This suggests that cysteine residue may be involved in the active site of the enzyme.

• Journal of Microbiology and Biotechnology
• /
• 제3권3호
• /
• pp.181-187
• /
• 1993

Steroid $\Delta^1$-dehydrogenase which introduces a double bond into the 1, 2 positions of steroid ring A was purified from Arthrobacter simplex, an excellent biotransformer of hydrocortisone into prednisolone. Hydrocortisone-induced cells were disrupted by vigorous agitation with glass beads, and a solubilized enzyme was obtained after centrifugation at 100, 000$\times$g for 90 minutes. The enzyme was purified 123-fold in three steps of chromatographic procedures with 13% yield. The last step of testosterone-agarose affinity column decisively contributed to the successful purification. The molecular weight of the enzyme was estimated to be 98, 000 by SDS-PAGE and 100, 000 by gel filtration, indicating that this enzyme behaves as a monomer. The enzyme showed demands for artificial electron acceptor, and among the several reagents tested, phenazine methosulfate acted as the most effective electron acceptor. Subcellular distribution of this enzyme was studied by centrifugation experiment. Comparison of the enzyme activities in pelleted membrane and cytosol fractions suggests that the enzyme may be a weakly attached peripheral membrane protein in vivo. But considerable amounts of enzyme was solubilized without any additional treatments for membrane protein.

• The Plant Pathology Journal
• /
• 제26권3호
• /
• pp.245-252
• /
• 2010

An endophytic bacterial strain, Arthrobacter humicola YC6002, was isolated from a surface sterilized root of Korean turf grass (Zoysia japonica) collected from Jinju, Korea. This strain showed inhibitory effect on germination and shoot growth of radish. The inhibition of germination and shoot growth of radish seeds varied depending on the age of culture and the temperature at which it was incubated. The culture filtrate of 1/10-strength Tryptic Soy Broth medium, incubated for 48 hours at $30^{\circ}C$, showed the highest inhibitory effect on radish seed germination and shoot growth (92% inhibition as compared to control). The active compound with seed germination and shoot growth inhibition was purified and identified as 3-phenylpropionic acid. The purified compound had 53% and 93% inhibitory effect on seed germination and shoot growth of radish for 500 and 1000 ppm solutions, respectively.

• 한국미생물·생명공학회지
• /
• 제32권4호
• /
• pp.366-370
• /
• 2004

Four chitinase producing bacteria, Arthrobacter nicotinae CH4, Arthrobacter nicotinae CHI3, Arthrobacter sp. CH5 and Micrococcus sp. CH3, were isolated from small crabs and shrimps. We investigated the optimum medium condition for the production of enzyme and high cell mass. The preferable medium composition was as follows: colchitin 0.1 %(w/v), glycerol 0.25%(w/v) and yeast extract 0.05%(w/v) in minimal midium ($K_{2}HPO_{4}$ 0.7 g/l, $KH_{2}PO_{4}$ 0.3 g/l, $MgSO_{4}{\cdot}5H_{2}O$ 0.5 g/l, $FeSO_{4}}{\cdot}7H_{2}O$ 0.01 g/l, $ZnSO_{4}$ 0.001 g/l, $MnCI_2$ 0.001 g/l, pH 7.0). This cell culture medium could be used directly as sample for measuring chitinase activity. Because it hardly conreducing sugar such as glucose (blank value=0), the detected reducing sugar can be considered as a chitinase reaction product. The results can be used for easy preparation method for determination of enzyme activity and analysis of enzyme-substrate reaction in step of screening of chitinase producing bacteria.

• Journal of Applied Biological Chemistry
• /
• 제42권2호
• /
• pp.71-74
• /
• 1999

The crude enzyme prepared from the culture supernantant of Arthrobacter sp. S37 was purified by Phenyl Toyopearl column chromatography. Six endo-inulinases were detected by activity staining on native PAGE and named Inu I to Inu VI. Endo-inulinase were further purified by DEAE cellulose column chromatography and band slicing. Inu II~VI produced mainly inulotriose (F3) and inulotetraose (F4) as well as a small amount of inulobiose (F2) and fructose in contrast to Inu I producing F3, F4 and F5 from inulin. The N-terminal amino acid sequence of native and six CNBr-cleaved fragment of Inu VI were determined. No homology was found in amino acid sequences between Inu VI and other fructan hydrolase including invertase reported.

• Food Science and Biotechnology
• /
• 제18권2호
• /
• pp.519-525
• /
• 2009

A bacterial strain (Strain N-08) capable of extracellularly producing high level of non-reducing oligosaccharide (NR-OS) isolated from soil. The strain was identified phylogenetically by 16S rDNA sequence analysis and found to be very close to Arthrobacter crystallopoietes. The high production of NR-OS was observed in the basal culture medium containing maltose as a sole carbon source. The NR-OS in culture supernatant was purified by glucoamylase treatment and Dowex-1 (OH.) ion exchange chromatography and its structure was characterized. This oligosaccharide consisted of only glucose. Methylation analysis indicated that this fraction was composed mainly of non-reducing terminal glucopyranoside. Matrixassisted laser-induced/ionization time-of-flight (MALDI-TOF) and electrospray ionization-mass spectrometry (ESI-MS)/MS analyses suggested that this oligosaccharide comprised non-reducing disaccharide unit with 1,1-glucosidic linkage. When this disaccharide was analyzed by $^1H$-NMR and $^{13}C$-NMR, it gave the same signals with $\alpha$-D-glucopyranosyl-(1,1)-$\alpha$-Dglucopyranoside. These results indicated that the NR-OS produced by A. crystallopoietes N-08 was ${\alpha}1$,${\alpha}1$-trehalose. This is the first report of the trehalose which can be produced directly from maltose by A. crystallopoietes N-08.