• Title/Summary/Keyword: Arthrobacter sp. characterization

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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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    • 제6권6호
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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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Isolation and Characterization of Nicotine-Degrading Bacterium Arthrobacter sp. NU11 and NU15 (니코틴 분해세균 Arthrobacter sp. NU11과 NU15의 분리 및 특성)

  • Jeong, Yeonju;Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • 제50권1호
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    • pp.67-72
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    • 2014
  • Minimal broth containing nicotine as a sole carbon source (MB/N) was used to isolate novel nicotine-degrading bacterial strains from tobacco plants and field soils. Comparative analysis of 16S rRNA gene sequence, phenotypic test and morphological tests showed that the position of these isolates were in the genus Arthrobacter of the family Micrococcaceae. The highest 16S rRNA gene sequence similarity of the isolate NU11 and NU15 to type strains in the genus Arthrobacter were Arthrobacter equi (98.2%) which was presumably a novel strain and Arthrobacter nicotinovorans (99.8%), respectively. Both strain NU11 and NU15 showed rod shaped, Gram-positive characteristics and catalase activity, but did not show oxidase activity. The novel strain NU11 was found to degrade efficiently nicotine in MB/N medium by the analysis of UV absorption spectra and could be used as an organism in bioremediation technique.

Characterization of PAH (Polycyclic Aromatic Hydrocarbon)-Degrading Bacteria Isolated from Commercial Gasoline (상용 휘발유로부터 분리한 다환 방향족 탄화수소(PAH) 분해 세균의 특성)

  • Kwon, Tae-Hyung;Woo, Jung-Hee;Park, Nyun-Ho;Kim, Jong-Shik
    • Korean Journal of Environmental Agriculture
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    • 제34권3호
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    • pp.244-251
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    • 2015
  • BACKGROUND: Recent studies have described the importance of bacteria that can degrade polycyclic aromatic hydrocarbons (PAHs). Here we screened bacterial isolates from commercial gasoline for PAH degraders and characterized their ability to degrade PAHs, lipids and proteins as well as their enantioselective epoxide hydrolase activity, salt tolerance, and seawater survival. METHODS AND RESULTS: One hundred two bacteria isolates from commercial gasoline were screened for PAH degraders by adding selected PAHs on to the surface of agar plates by the sublimation method. A clear zone was found only around the colonies of PAH degraders, which accounted for 13 isolates. These were identified as belonging to Bacillus sp., Brevibacterium sp., Micrococcus sp., Corynebacterium sp., Arthrobacter sp., and Gordonia sp. based on 16S rRNA sequences. Six isolates belonging to Corynebacterium sp., 3 of Micrococcus sp., Arthrobacter sp. S49, and Gordonia sp. H37 were lipid degraders. Arthrobacter sp. S49 was the only isolate showing high proteolytic activity. Among the PAH-degrading bacteria, Arthrobacter sp. S49, Brevibacterium sp. S47, Corynebacterium sp. SK20, and Gordonia sp. H37 showed enantioselective epoxide hydrolase activity with biocatalytic resolution of racemic styrene oxide. Among these, highest enantioselective hydrolysis activity was seen in Gordonia sp. H37. An intrinsic resistance to kanamycin was observed in most of the isolates and Corynebacterium sp. SK20 showed resistance to additional antibiotics such as tetracycline, ampicillin, and penicillin. CONCLUSION: Of the 13 PAH-degraders isolated from commercial gasoline, Arthrobacter sp. S49 showed the highest lipid and protein degrading activity along with high active epoxide hydrolase activity, which was the highest in Gordonia sp. H37. Our results suggest that bacteria from commercial gasoline may have the potential to degrade PAHs, lipids, and proteins, and may possess enantioselective epoxide hydrolase activity, high salt tolerance, and growth potential in seawater.

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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    • 제26권8호
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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.

Isolation and Characterization of Endo-inulinases from Arthrobacter sp. S37

  • Koo, Bong-Seong;Kang, Su-Il;Kim, Su-Il
    • Journal of Applied Biological Chemistry
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    • 제42권2호
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    • pp.71-74
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    • 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.

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Isolation and Characterization of 3,4-Dichloroaniline Degrading Bacteria from a Sandbank (갯벌에서 분리한 3,4-Dichloroaniline 분해 미생물의 특성)

  • Kim, Young-Mog
    • Journal of Marine Bioscience and Biotechnology
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    • 제1권4호
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    • pp.275-281
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    • 2006
  • The compound 3,4-dichloroaniline (DCA) is an aromatic amine used as an intermediate product in the synthesis of herbicides, azo-dyes and harmaceuticals. It is also a degradation product of some herbicides (diuron, propanil, and linuron) and of trichlorocarbanilide, a chemical used as active agent in the cosmetic industry. 3,4-DCA, however, is considered potential pollutants due to their toxic and recalcitrant properties to humans and other species. A bacterium capable of growth on 3,4-DCA was isolated by dilution method from 3,4-DCA-containing enrichment culture. Finally, a strain, YM-14, capable of degrading efficiently 3,4-DCA was isolated from a sandbank. The isolated strain, YM-14 was identified to be Arthrobacter sp.. Fifty ppm 3,4-DCA in 1/10 LB media was completely degraded by the growth of Arthrobacter sp. YM-14 for 12 h at $30^{\circ}C$. The isolated strain is capable of growth on 3,4-DCA as sole carbon source and also able to degrade other chloroaniline compounds. Also, the isolated strain showed high level of catechol 1,2-dioxygenase activity by 3,4-DCA exposure. The catechol 1,2-dioxygenase was supposed to be ones of the important factors for 3,4-DCA degradation.

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Molecular Characterization of Cold-Inducible ${\beta}$-Galactosidase from Arthrobacter sp. ON14 Isolated from Antarctica

  • Xu, Ke;Tang, Xixiang;Gai, Yingbao;Mehmood, Muhammad Aamer;Xiao, Xiang;Wang, Fengping
    • Journal of Microbiology and Biotechnology
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    • 제21권3호
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    • pp.236-242
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    • 2011
  • A psychrotrophic bacterium, Arthrobacter sp. ON14, isolated from Antarctica, was shown to exhibit a high ${\beta}$-galactosidase activity at a low temperature. A genomic library of ON14 was constructed and screened for ${\beta}$-galactosidase genes on functional plates containing 5-bromo-4-chloro-3-indolyl-${\beta}$-D-galactopyranoside (X-gal) as the substrate. Two different ${\beta}$-galactosidase genes, named as galA, galB, were found in ON14. Computational analyses of the genes revealed that the encoded protein GalA belongs to family 2 of glycosyl hydrolysases and is a cold-active protein, whereas GalB belongs to family 42 of glycosyl hydrolysases and is a mesophilic protein. Reverse transcription analyses revealed that the expression of galA is highly induced at a low temperature ($4^{\circ}C$ ) and repressed at a high temperature ($28^{\circ}C$ ) when lactose is used as the sole carbon source. Conversely, the expression of galB is inhibited at a low temperature and induced at a high temperature. The purified GalA showed its peak activity at $15^{\circ}C$ and pH 8. The mineral ions $Na^+$, $K^+$, $Mg^{2+}$, and $Mn^{2+}$ were identified as enzyme activators, whereas $Ca^{2+}$ had no influence on the enzyme activity. An enzyme stability assay revealed that the activity of GalA is significantly decreased when it is incubated at $45^{\circ}C$ for 2 h, and all its activity is lost when it is incubated at $50^{\circ}C$.

Suppression of Fusarium Wilt Caused by Fusarium oxysporum f. sp. lactucae and Growth Promotion on Lettuce Using Bacterial Isolates

  • Yadav, Dil Raj;Adhikari, Mahesh;Kim, Sang Woo;Kim, Hyun Seung;Lee, Youn Su
    • Journal of Microbiology and Biotechnology
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    • 제31권9호
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    • pp.1241-1255
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    • 2021
  • This study was carried out to explore a non-chemical strategy for enhancing productivity by employing some antagonistic rhizobacteria. One hundred eighteen bacterial isolates were obtained from the rhizospheric zone of various crop fields of Gangwon-do, Korea, and screened for antifungal activity against Fusarium wilt (Fusarium oxysporum f. sp. lactucae) in lettuce crop under in vitro and in vivo conditions. In broth-based dual culture assay, fourteen bacterial isolates showed significant inhibition of mycelial growth of F. oxysporium f. sp. lactucae. All of the antagonistic isolates were further characterized for the antagonistic traits under in vitro conditions. The isolates were identified on the basis of biochemical characteristics and confirmed at their species level by 16S rRNA gene sequencing analysis. Arthrobacter sulfonivorans, Bacillus siamensis, Bacillus amyloliquefaciens, Pseudomonas proteolytica, four Paenibacillus peoriae strains, and Bacillus subtilis were identified from the biochemical characterization and 16S rRNA gene sequencing analysis. The isolates EN21 and EN23 showed significant decrease in disease severity on lettuce compared to infected control and other bacterial treatments under greenhouse conditions. Two bacterial isolates, EN4 and EN21, were evaluated to assess their disease reduction and growth promotion in lettuce in field conditions. The consortium of EN4 and EN21 showed significant enhancement of growth on lettuce by suppressing disease caused by F. oxysporum f. sp. lactucae respectively. This study clearly indicates that the promising isolates, EN4 (P. proteolytica) and EN21 (Bacillus siamensis), can be commercialized and used as biofertilizer and/or biopesticide for sustainable crop production.

Cloning, Expression, and Characterization of Bacillus sp. snu-7 Inulin Fructotransferase

  • Kim, Chung-Sei;Hong, Chang-Ki;Kim, Kyoung-Yun;Wang, Xiu-Ling;Kang, Su-Il;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • 제17권1호
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    • pp.37-43
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    • 2007
  • A gene encoding inulin fructotransferase (di-D-fructofuranose 1,2': 2,3' dianhydride [DFA III]-producing IFTase, EC 4.2.2.18) from Bacillus sp. snu-7 was cloned. This gene was composed of a single, 1,353-bp open reading frame encoding a protein composed of a 40-amino acid signal peptide and a 410-amino acid mature protein. The deduced amino acid sequence was 98% identical to Arthrobacter globiformis C11-1 IFTase (DFA III-producing). The enzyme was successfully expressed in E. coli as a functionally active, His-tagged protein, and it was purified in a single step using immobilized metal affinity chromatography. The purified enzyme showed much higher specific activity (1,276 units/mg protein) than other DFA III-producing IFTases. The recombinant and native enzymes were optimally active in very similar pH and temperature conditions. With a 103-min half-life at $60^{\circ}C$, the recombinant enzyme was as stable as the native enzyme. Acidic residues and cysteines potentially involved in the catalytic mechanism are proposed based on an alignment with other IFTases and a DFA IIIase.