• Title/Summary/Keyword: Arthrobacter sp. A-6

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Production of Inulin fructotransferase(depolumerizing)by Arthrobacter sp. A-6 (Arthrobacter sp. A-6에 의한 Inulin Fructotransferase (depolymerizing)의 생산)

  • 박정복;권영만;최용진
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.68-74
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    • 1995
  • A bacterial strain A-6 producing the high level of an extracellular inulin fructotransfe rase(depolymerizing)(EC 2.4.1.93) which converts inulin into di-D-fructofuranose dianhydride III (DFAIII) was isolated from soil. The isolated strain could be classified as a species belonging to the genus Arthrobacter based on its morphological and physiological characteristics identified in this work. Production of the enzyme was induced by inulin, and the highest activity was detected in the slightly acidic medium supplemented with 2.5% inulin and 0.1% trypton as a sole carbon and a nitrogen source, respectively. Under the optimal conditions, the enzyme activity in the culture supernatant reached approximately 60 uints/ml after 96 hours of cultivation. The optimum pH and temperature for the crude enzyme preparation from Arthrobacter sp. A-6 were pH 5.0 and 60$\circ$C , respectively. The DFA produced by the action of the inulin fructotransferase was confirmed to be DFAIII by paper chromatography, HPLC and $^{13}$C-NMR spectroscopy.

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Purification and Characterization of Inulin Fructotransferase (Depolymerizing) from Arthrobacter sp. A-6

  • PARK, JEONG-BOK;YONG-JIN CHOI
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.402-406
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    • 1996
  • Inulin fructotransferase (depolymerizing) (EC 2.4.1.93) was purified 34-fold from the culture broth of Arthrobacter sp. A-6 by using a combination of ammonium sulfate fractionation, DEAE-Sepharose CL-6B chromatography and Sephacryl S-200 gel filtration. The purified enzyme converts inulin into di-D-fructofuranose dianhydride III(DFA III) and small quantities of fructo-oligosaccharides. The temperature and pH optima of the enzyme were $70^{\circ}C$ and 6.0, respectively. Molecular weight of the enzyme was determined to be 49 kDa by 12$%$ SDS-polyacrylamide gel electrophoresis, and 145 kDa by Sephacryl S-200gel filtration. This indicates that the functional inulin fructotransferase of Arthrobacter sp. A-6 has a homomeric trimer structure. The enzyme had an isoelectric point of pH 4.6. The N-terminal amino acid sequence of the purified enzyme subunit was Ala-Asp-Asn-Pro-Asp-Gly(\ulcorner)-Ser-Asn-Met(or Glu)-Tyr-Asp-Val.

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Cloning and Expression of Inulin Fructotransferase Gene of Arthrobacter sp. A-6 in Escherichia coli and Bacillus subtilis

  • Kim, Hwa-Young;Kim, Chan-Wha;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.275-280
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    • 2000
  • The inulin fructotransferse (depolymerizing) (IFTase, EC 2.4.1.93) gene of Arthrobacter sp. A-6 was cloned and expressed in Escherichia coli and Bacillus subtilis. The IFTase gene consisted of an ORF of 1.311 nucleotides encoding a polypeptide of 436 amino acids containing a signal peptide of 31 amino acids in the N-terminus. The molecular mass of the IFTase based on the nucleotide sequence was calculated to be 46.116 Da. The recombinant E. coli $DH5{\alpha}$ cells expressing the Arthrobacter sp. A-6 IFTase gene produced most of the IFTase intracelularly. In contrast, the recombinant B. subtilis DB 104 carrying the IFTas gene on a B. subtilis-E. Coli expression vector secreted the IFTase into the culture fluid efficiently.

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Isolation and Characteristics of Exopolysaccharide Producing Bacteria in a Ginseng Root System (인삼 근계로부터 다당 생성세균의 분리 및 특성)

  • Cho, Geon-Yeong;Jeon, In-Hwa;Han, Song-Ih;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.297-300
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    • 2013
  • EPS producing bacteria were enumerated in ginseng root system (rhizosphere soil, rhizoplane, inside of root). EPS producing bacterial density of rhizosphere soil, rhizoplane and inside of root were distributed $9.0{\times}10^6$ CFU/g, $7.0{\times}10^6$ CFU/g, and $1.4{\times}10^3$ CFU/g, respectively. Phylogenetic analysis of the 24 EPS producing isolates based on the 16S rRNA gene sequences, EPS producing isolates from rhizosphere soil (RS) belong to genus Arthrobacter (6 strains) and Rhizobium (1 strain). EPS producing bacteria from rhizoplane (RP) were Arthrobacter (6 strains), Rhodococcus (1 strain) and Pseudomonas (1 strain). EPS producing bacteria from inside of root (IR) were categorized into Rhzobium (6 strains), Bacillus (1 strain), Rhodococcus (1 strain), and Pseudomonas (1 strain). Phylogenetic analysis indicated that Arthrobacter may be a member of representative EPS producing bacteria from ginseng rhizosphere soil and rhizoplane, and Rhizobium is typical EPS producing isolates from inside of ginseng root. The yield of EPS was 10.0 and 4.9 g/L by Rhizobium sp. 1NP2 (KACC 17637) and Arthrobacter sp. 5MP1 (KACC 17636). The purified EPS were analyzed by Bio-LC and glucose, galactose, mannose and glucosamine were detected. The major EPS sugar of these strains was glucose (72.7-84.9%).

Isolation of Indole-3-acetic acid (IAA) producing Arthrobacter sp. and plant growth promotion effect (Indole-3-acetic acid (IAA) 생성 Arthrobacter sp.의 분리 및 식물 생육촉진 효과)

  • Da Som Kim;Ho-Young Shin;Song-Ih Han
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.6
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    • pp.831-838
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    • 2022
  • An auxin-producing bacteria, KSD16, KSD33, and KSD36 were isolated from agricultural soil. The strain KSD16, KSD33, and KSD36 was classified as a strain of Arthrobacter sp. based on phylogenetic analysis of 16S rRNA gene. The isolated KDS16, KDS33, and KSD36 was confirmed to produce indole-3-acetic acid (IAA), which is one of the auxin hormones. When the concentration of IAA was assessed the maximum concentration of IAA, 206.62 mg L-1, was detected from the culture broth incubated in R2A medium containing 0.1% L-tryptophan for 48 h at 28 ℃. To study the effect of IAA producing bacteria on germination rate, seeds of Mung bean were prepared for each treatment. KSD16, KSD33, and KSD36 showed significant increase in root length and number of adventitious roots than the controls. To investigate the growth-promoting effects on the crops, Arthrobacter species were placed in water cultures and seed pots of mung beans. In consequence, the seed germination of mung beans was 73.4% higher than the control.

Optimization of Physical Factor for amylase Production by Arthrobacter sp. by Response Surface Methodology (반응표면분석법을 통한 Arthrobacter sp.의 amylase 생산 최적화)

  • Kim, Hyun-do;Im, Young-kum;Choi, Jong-il;Han, Se Jong
    • Korean Chemical Engineering Research
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    • v.54 no.1
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    • pp.140-144
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    • 2016
  • In this study, the physical factors for amylase production by Arthrobacter sp. were optimized using response surface methodology(RSM). Antarctic microorganism Arthrobacter sp. PAMC 27388 was obtained from the Polar and Alpine Microbial Collection(PAMC) at the Korea Polar Research Institute. This microorganism was confirmed for the excretion of amylase with Lugol's solution. The amylase activity was after flask culture was as low as 1.66 mU/L before optimization. The physical factors including the inoculum volume, the initial culture pH, and the medium volume were chosen to be optimized for the enhanced amylase production. The calculated results using RSM indicate that the optimal physical factors were 2.49 mL inoculum volume, 6.85 pH and 42.87 mL medium volume with a predicted amylase production of 2.84 mU/L. The experimentally obtained amylase activity was 2.50 mU/L, which was a 150% increase compared to the level before optimization.

Optimization of Medium for Carotenoids Production by Arthrobacter sp. PAMC 25486 Using Response Surface Methodology (반응표면분석법을 이용한 Arthrobacter sp. PAMC 25486의 카로티노이드 생산배지 최적화)

  • Kim, Hyun-Do;Choi, Jong-Il;Han, Se Jong
    • Korean Chemical Engineering Research
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    • v.52 no.6
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    • pp.834-839
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    • 2014
  • This study was conducted to optimize the medium composition for carotenoid production in Arthrobacter sp. PAMC 25486 through response surface methodology (RSM). Using a Placket-Burman design, from which yeast extract, $MgSO_4$ and dextrose were identified as the significant factors affecting carotenoids production. RSM studies for carotenoids production by Arthrobacter sp. PAMC 25486 have been carried out for three parameters of yeast extract, $MgSO_4$ and dextrose concentrations. These significant factors were optimized by experiments and RSM, as 1 g/L yeast extract, 0.0879 g/L $MgSO_4$ and 1 g/L dextrose. The experimentally obtained concentration of carotenoid was 288 mg/L, and it became 2-fold increase on concentration before optimization.

Isolation of Extracellular Cytosine Deaminase Producing Strain Arthrobacter sp. JH-13 and Cultural Conditions of It's Enzyme Production (세포의 Cytosine Deaminase 생산균 Arthrobacter sp. JH-13의 분리 및 효소생산 조건)

  • 전홍기;박정혜
    • Korean Journal of Microbiology
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    • v.22 no.4
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    • pp.257-263
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    • 1984
  • A strain producing an extracellular cytosine deaminase was isolated from soil samples. The enzyme obtained from the strain possessed the substrate specificity to both cytosine and 5-fluorocytosine. From the results of its morphological, cultural, physiological, and biochemical properties, the strain was thought to be the genus Arthrobacter. Therefore, it was named as Arthrobacter sp. JH-13. The composition of optimum medium for the enzyme formation was 0.5% of peptone, 0.5% of meat extract, 0.5% of soluble starch, and 0.1% of KCl. The optimum pH for the enzyme formation was 8.0. When the microoganism was cultured aerobically in the above medium, enzyme production reached at maximum in 54 hours at $30^{\circ}C$.

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Biosorption of Lead(II) by Arthrobacter sp. 25: Process Optimization and Mechanism

  • Jin, Yu;Wang, Xin;Zang, Tingting;Hu, Yang;Hu, Xiaojing;Ren, Guangming;Xu, Xiuhong;Qu, Juanjuan
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1428-1438
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    • 2016
  • In the present work, Arthrobacter sp. 25, a lead-tolerant bacterium, was assayed to remove lead(II) from aqueous solution. The biosorption process was optimized by response surface methodology (RSM) based on the Box-Behnken design. The relationships between dependent and independent variables were quantitatively determined by second-order polynomial equation and 3D response surface plots. The biosorption mechanism was explored by characterization of the biosorbent before and after biosorption using atomic force microscopy (AFM), scanning electron microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. The results showed that the maximum adsorption capacity of 9.6 mg/g was obtained at the initial lead ion concentration of 108.79 mg/l, pH value of 5.75, and biosorbent dosage of 9.9 g/l (fresh weight), which was close to the theoretically expected value of 9.88 mg/g. Arthrobacter sp. 25 is an ellipsoidal-shaped bacterium covered with extracellular polymeric substances. The biosorption mechanism involved physical adsorption and microprecipitation as well as ion exchange, and functional groups such as phosphoryl, hydroxyl, amino, amide, carbonyl, and phosphate groups played vital roles in adsorption. The results indicate that Arthrobacter sp. 25 may be potentially used as a biosorbent for low-concentration lead(II) removal from wastewater.

Cultivation of Arthrobactor sp. A-6 and Production of DFA III(Di-Fructofuranose Dianhydride) from Chicory Root Extract (Arthrobactor sp. A-6의 배양과 Chicory 뿌리 추출물에서 Di-Fructofuranose Dianhydride(DFAIII)의 생산)

  • 김기은;신창훈;최용진;김찬화
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.69-73
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    • 2000
  • Arthrobacter sp. A-6 was cultivated and DFA III(di-fructofuranose dianhydride) was produced with inulin fructotransferase from the chicory root. The specific growth rate, yield of cell mass and yield of enzyme from the culture in variable chicory root extracts were studied and the results compared. Standard inulin solution(10%) was treated with the crude enzyme solution of inulin fructotransferase from the cell culture, 1.14mg/ml of DFA III was produced. The enzyme reactions were processed with various preparations of chicory root extracts in the same conditions. The highest yield of DFA III production(2.29 mg/ml) was obtained from the chicory roots without washing or extraction. The yield of DFA III from the washed chicory roots without extraction was at lowest(0.44 mg/ml). The production process of inulin fructotransferase and DFA III from the chicory root without prewashing or extraction steps were more efficient.

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