• Title/Summary/Keyword: $sp^{3}$ carbon

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Induction by Carvone of the Polychlorinated Biphenyl (PCB)-Degradative Pathway in Alcaligenes eutrophus H850 and Its Molecular Monitoring

  • Park, Young-In;So, Jae-Seong;Koh, Sung-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.804-810
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    • 1999
  • There is a possibility that carvone, a monoterpene from spearmint (Mentha spicata), could induce the bph degradative pathway and genes in Alcaligenes eutrophus H850, which is a known Gram-negative PCB degrader with a broad substrate specificity that was thoroughly investigated with Arthrobacter sp. BIB, a Gram-positive PCB degrader. The strains BIB and H850 were unable to utilize and grow on the plant terpene [(R)-(-)-carvone] (50ppm) to be recognized as a sole carbon source. Nevertheless, the carvone did induce 2,3-dihydroxybiphenyl 1,2-dioxygenase (encoded by bphC) in the strain B lB, as observed by a resting cell assay that monitors accumulation of a yellow meta ring fission product from 4,4'-dichlorobiphenyl (DCBp). The monoterpene, however, did not appear to induce the meta cleavage pathway in the strain H850. Instead, an assumption was made that the strain might be using an alternative pathway, probably the ortho-cleavage pathway. A reverse transcription (RT)-PCR system, utilizing primers designed from a conserved region of the bphC gene of Arthrobacter sp. M5, was employed to verify the occurrence of the alternative pathway. A successful amplification (182bp) of mRNA transcribed from the N-terminal region of the bphC gene was accomplished in H850 cells induced by carvone (50ppm) as well as in biphenyl-growth cells. It is, therefore, likely that H850 possesses a specific PCB degradation pathway and hence a different substrate specificity compared with B1B. This study will contribute to an elucidation of the dynamic aspects of PCB bioremediation in terms of roles played by PCB degraders and plant terpenes as natural inducer substrates that are ubiquitous and environmentally compatible.

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Arthrobacter sp. Strain KU001 Isolated from a Thai Soil Degrades Atrazine in the Presence of Inorganic Nitrogen Sources

  • Sajjaphan, Kannika;Heepngoen, Pimpak;Sadowsky, Michael J.;Boonkerd, Nantakorn
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.602-608
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    • 2010
  • An atrazine-degrading bacterium, strain KU001, was obtained from a sugarcane field at the Cane and Sugar Research and Development Center at the Kasetsart University, Kamphaeng Saen Campus, Thailand. Strain KU001 had a rod-to-coccus morphological cycle during growth. Biolog carbon source analysis indicated that the isolated bacterium was Arthrobacter histidinolovorans. Sequence analysis of the PCR product indicated that the 16S rRNA gene in strain KU001 was 99% identical to the same region in Arthrobacter sp. The atrazine degradation pathway in strain KU001 consisted of the catabolic genes trzN, atzB, and atzC. Strain KU001 was able to use atrazine as a sole nitrogen source for growth, and surprisingly, atrazine degradation was not inhibited in cells grown on ammonium, nitrate, or urea, as compared with cells cultivated on growth-limiting nitrogen sources. During the atrazine degradation process, the supplementation of nitrate completely inhibited atrazine degradation activity in strain KU001, whereas ammonium and urea had no effect on atrazine degradation activity. The addition of strain KU001 to sterile or nonsterile soils resulted in the disappearance of atrazine at a rate that was 4- to 5-fold more than that achieved by the indigenous microbial community. The addition of citrate to soils resulted in enhanced atrazine degradation, where 80% of atrazine disappeared within one day following nutrient supplementation.

Properties and Functions of Melanin Pigment from Klebsiella sp. GSK

  • Sajjan, Shrishailnath S.;Anjaneya, O;Kulkarni, Guruprasad B.;Nayak, Anand S.;Mashetty, Suresh B.;Karegoudar, T.B.
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.60-69
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    • 2013
  • Purified melanin pigment from Klebsiella sp. GSK was characterized by thermogravimetric, differential thermal, X-ray diffraction and elemental analysis. This melanin pigment is structurally amorphous in nature. It is thermally stable up to $300^{\circ}C$ and emits a strong exothermic peak at $700^{\circ}C$. Its carbon, hydrogen and nitrogen composition is 47.9%, 6.9% and 12.0%, respectively. It was used to scavenge metal ions and free radicals. After immobilizing the pigment and using it to adsorb copper and lead ions, the metal ion adsorption capacity was evaluated by atomic absorption spectroscopy (AAS) and the identity of melanin functional groups involved in the binding of metal ions was determined by Fourier transform infrared (FT-IR) spectroscopy. Batch adsorption studies showed that 169 mg/g of copper and 280 mg/g of lead were adsorbed onto melanin-alginate beads. The metal ion adsorption capacity of the melanin-alginate beads was relatively significant compared to alginate beads. The metal ion desorption capacity of HCl was greater (81.5% and 99% for copper and lead, respectively) than that of EDTA (80% and 71% for copper and lead, respectively). The ability of the melanin pigment to scavenge free radicals was evaluated by inhibition of the oxidation of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and was shown to be about 74% and 98%, respectively, compared with standard antioxidants.

Activation and immobilization of phenol-degrading bacteria on oil palm residues for enhancing phenols degradation in treated palm oil mill effluent

  • Tosu, Panida;Luepromchai, Ekawan;Suttinun, Oramas
    • Environmental Engineering Research
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    • v.20 no.2
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    • pp.141-148
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    • 2015
  • The presence of phenols in treated palm oil mill effluent (POME) is an environmental concern due to their phytotoxicity and antimicrobial activity. In this study, phenol-degrading bacteria, Methylobacterium sp. NP3 and Acinetobacter sp. PK1 were immobilized on oil palm empty fruit bunches (EFBs) for removal of phenols in the treated POME. The bacterial exopolysaccharides (EPS) were responsible for cell adhesion to the EFBs during the immobilization process. These immobilized bacteria could effectively remove up to 5,000 mg/L phenol in a carbon free mineral medium (CFMM) with a greater degradation efficiency and rate than that with suspended bacteria. To increase the efficiency of the immobilized bacteria, three approaches, namely activation, acclimation, and combined activation and acclimation were applied. The most convenient and efficient strategy was found when the immobilized bacteria were activated in a CFMM containing phenol for 24 h before biotreatment of the treated POME. These activated immobilized bacteria were able to remove about 63.4% of 33 mg/L phenols in the treated POME, while non-activated and/or acclimated immobilized bacteria could degrade only 35.0%. The activated immobilized bacteria could be effectively reused for at least ten application cycles and stored for 4 weeks at $4^{\circ}C$ with the similar activities. In addition, the utilization of the abundant EFBs gives value-added to the palm oil mill wastes and is environmentally friendly thus making it is attractive for practical application.

Effect of Substrate Bias Voltage on DLC Films Prepared by ECR-PECVD (ECR-PECVD 방법으로 제작된 DLC 박막의 기판 Bias 전압 효과)

  • 손영호;정우철;정재인;박노길;김인수;김기홍;배인호
    • Journal of the Korean Vacuum Society
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    • v.9 no.4
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    • pp.328-334
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    • 2000
  • DLC (Diamond-Like Carbon) films were deposited by ECR-PECVD (electron cyclotron resonance plasma-enhanced chemical vapor deposition) method with the variation of substrate bias voltage under the others are constant except it. We have investigated the ion bombardment effect induced by the substrate bias voltage on films during the deposition of film. The characteristics of the film were analyzed using the Dektak surface profiler, SEM, FTIR spectroscopy, Raman spectroscopy and Nano Indentation tester. FTIR spectroscopy analysis shows that the amount of dehydrogenation in films was increased with the increase of substrate bias voltage and films thickness was decreased. Raman scattering analysis shows that integrated intensity ratio $(I_D /I_G)$ of the D and G peak was increased as the substrate bias voltage increased, and films hardness was increased. From these results, it can be concluded that films deposited at this experimental have the enhanced characteristics of DLC because of the ion bombardment effect on films during the deposition of film.

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Decolorization and Biotransformation of Triphenylmethane Dye, Methyl Violet, by Aspergillus sp. Isolated from Ladakh, India

  • Kumar, C. Ganesh;Mongolla, Poornima;Basha, Anver;Joseph, Joveeta;Sarma, V.U.M.;Kamal, Ahmed
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.267-273
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    • 2011
  • Methyl violet, used extensively in the commercial textile industry and as a biological stain, is a hazardous recalcitrant. Aspergillus sp. strain CB-TKL-1 isolated from a water sample from Tsumoriri Lake, Karzok, Ladakh, India, was found to completely decolorize methyl violet within 24 h when cultured under aerobic conditions at $25^{\circ}C$. The rate of decolorization was determined by monitoring the decrease in the absorbance maxima of the dye by UV-visible spectroscopy. The decolorization of methyl violet was optimal at pH 5.5 and $30^{\circ}C$ when agitated at 200 rpm. Addition of glucose or arabinose (2%) as a carbon source and sodium nitrate or soyapeptone (0.2%) as a nitrogen source enhanced the decolorization ability of the culture. Furthermore, the culture exhibited a maximum decolorization rate of methyl violet after 24 h when the C:N ratio was 10. Nine N-demethylated decolorized products of methyl violet were identified based on UV-visible spectroscopy, Fourier transform infrared (FTIR), and LC-MS analyses. The decolorization of methyl violet at the end of 24 h generated mono-, di-, tri-, tetra-, penta-, and hexa-N-demethylated intermediates of pararosaniline. The variation of the relative absorption peaks in the decolorized sample indicated a linear decrease of hexa-N-demethylated compounds to non-N-demethylated pararosaniline, indicating a stepwise N-demethylation in the decolorization process.

Production and Characterization of Extracellular Phospholipase D from Streptomyces sp. YU100

  • Lim, Si-Kyu;Choi, Jae-Woong;Chung, Min-Ho;Lee, Eun-Tae;Khang, Yong-Ho;Kim, Sang-Dal;Nam, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.189-195
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    • 2002
  • Using Streptomyces sp. YU100 isolated from Korean soil, the fermentative production of phospholipase D was attempted along with its purification and characterization studies. When different carbon and nitrogen sources were supplemented in the culture medium, glucose and yeast extract were found to be the best. By varying the concentration of nutrients and calcium carbonate, the optimal culture medium was determined as 2.0% glucose, 1.5% yeast extract, 0.5% tryptone 0.3% calcium carbonate. During cultivation, the strain secreted most of the phospholipase D in the early stage of growth within 24 h. The phospholipase D produced in the culture broth exhibited hydrolytic activity as well as transphosphatidylation activity on lecithin (phosphatidylcholine). In particular, the culture broth showed 8.7 units/ml of hydrolytic activity when cultivated at $28^{\circ}C$ for 1.5 days. The phospholipase D was purified using 80% ammonium sulfate precipitation and DEAE-Sepharose CL-6B column chromatography, which produced a major band of 57 kDa on a 10% SDS-polyacrylamide gel with purity higher than 80%. The enzyme showed an optimal pH of 7 in hydrolytic reaction, and at pH 4 in a transphosphatidylation reaction. The enzyme activity increased until the reaction temperature was elevated to $60^{\circ}C$. The enzyme was relatively stable at high temperatures and neutral pH, but significantly unstable in the alkaline range. Among the detergents tested as emulsifiers of phospholipids, the highest enzyme activity was observed when 1.5% Triton X-100 was employed. However, no inhibitory effect by metal ions was detected. Under optimized reaction conditions, the purified enzyme not only completely decomposed PC to phosphatidic acid within 1 h, but also exhibited higher than 80% conversion rate of PC to PS by transphosphatidylation within 4 h.

Isolation of Fibrinolytic Yeasts from Korean Traditional Fermented Soybean (전통장류로부터 혈전용해 활성이 우수한 효모균주의 분리)

  • Yi, Jae-Hyoung;Heo, Nam-Kee;Choi, Byung-Gon;Park, Eun-Hee;Kwun, Se-Young;Kim, Myoung-Dong;Hong, Wun-Pyo;Yeo, Soo-Hwan;Baek, Seong-Yeol
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.184-189
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    • 2014
  • Yeast strains demonstrating fibrinolytic activity were isolated from traditional fermented soybean in Gangwon province, Korea. The AFY-1 strain isolated from fermented soybean paste showed the highest fibrinolytic activity (3.5 U/mg protein) corresponding to a 1.75 fold higher fibrinolytic activity compared with the plasmin (2.0 U/mg protein). The optimum temperature for the growth of AFY-1 strain was $32^{\circ}C$. Analysis of 18S rRNA gene sequence and carbon source utilization pattern indicated that the AFY-1 strain shares the highest homology (99%) with Saccharomycetales sp.

Investigation of Structural Change of DLC Coating during Frictional Wear by Raman Spectroscopy (라만 분광법을 활용한 마모 중 DLC 코팅의 구조적 변화 조사)

  • Kim, Song-Hee;Jang, Jae Cheol
    • Journal of the Korean institute of surface engineering
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    • v.52 no.1
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    • pp.16-22
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    • 2019
  • The structural change of DLC coatings during long-term wear test and dicing test under the low loading condition was investigated. DLC coatings were applied for the precision injection molds of a modified SNCM steel for the extension of life and the micro-diamond blades for the high cutting efficiency and the increase in life. A ball-on-disc wear tests in the mold steel and a dicing tests in the micro-diamond blades were conducted to understand degradation of DLC coatings. The degradation of DLC coatings for the injection mold steel and the micro-diamond blades during the wear and dicing tests were studied with Raman Spectroscopy. Raman peaks were divided two bands(D band and G band) to study the degradation process of DLC structure. By the wear test, polished condition of wear marks were observed to be maintained until 10 hrs of wear test period is given, but small striation marks appeared in 20 hours wear test. It was observed that $I_D/I_G$ ratios changed as the degradation of DLC coatings is proceeded during the wear tests and the dicing tests. It is suggested that the change in $I_D/I_G$ value possibly reflected from the composition of $sp^2$ and $sp^3$ bondings in DLC layers relevant to the change in mechanical and physical property.

Isolation of Biopolymer-producing Bacterium and Its Growth Pattern (Biopolymer 생산세균의 분리 및 증식패턴)

  • Yoo, Jin-Young;Chung, Dong-Hyo
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.421-428
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    • 1989
  • A soil bacterium synthesizing an extremely viscous biopolymer was isolated and identified as Pseudomonas delafieldii. The optimal pH and temperature for the growth were 6.5 and 3$0^{\circ}C$, respectively. Maximum specific growth rate was 0.24 h$^{-1}$. The specific polysaccharide productivity, growth yield and product yield were 6.25 mg/g-cell/h, 54.5% and 38.39%, respectively. The polysaccharide was presumed to be $\beta$-glucan containing glucose and gluconolactone (1.9:1.0 in molar ratio) and 1.35 % acetyl group, Element analysis showed that it contained carbon (31.85%) and hydrogen (5.15%). The weight average molecular weight by GPC was 5.64$\times$10$^7$. The intrinsic viscosity was 42.84 dl/g.

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