• Applied Biological Chemistry
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• 제30권3호
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• pp.219-226
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• 1987

Peroxidase isozyme C was isolated from mung bean cotyledon and purified to homogeneity as ascertained by chromatography and polyacrylamide gel electrophoresis, and then crystallized. Purification procedures included ammonium sulfate precipitation and column chromatography on Sephadex G-75, DEAE-cellulose and DEAE-Sephadex A-50. Peroxidase isozyme C was purified about 63 fold with 5% recovery. Isozyme C showed optimal activity at pH 5.0 with o-dianisidine and at pH 6.0 with guaiacol as substrate, and the optimal temperature was $70^{\circ}C$. Molecular weight of 50,000 was estimated for the isozyme C by SDS-polyacrylamide gel electrophoresis. At $70^{\circ}C$, it took 30 min to inactivate the isozyme to 50%, and at $80^{\circ}C$, this isozyme was almost completely inactivated in 20 min. The Km value of isozyme C for o-dianisidine was 0.11mM and that for guaiacol was 60.98mM using hydrogen peroxide as cosubstrate, and the kinetic pattern showed a competitive cyanide inhibition with respect to substrate. The crystalline structure of isozyme C was rectangular in shape.

• Asian-Australasian Journal of Animal Sciences
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• 제20권9호
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• pp.1424-1432
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• 2007

Two male, rumen fistulated crossbred Brahman-Thai native beef cattle (body weight = $400{\pm}50$ kg), fed on rice straw as a source of roughage, were used as rumen fluid sources. The treatments were $2{\times}3$ factorial arrangements; two roughages (fresh cassava foliage and cassava hay) and three sulfur levels (elemental sulfur) at 0.2 (control), 0.5 and 1% of DM, respectively. The experiment revealed that the rates (c) of gas production, ammonia-nitrogen concentration, true digestibility, total concentration or molar proportions of VFA and microbial biomass were not significantly different between cassava hay and fresh cassava foliage. However, all parameters for cassava hay were higher than for fresh cassava foliage. The supplementation of 0.5% sulfur to fresh cassava foliage resulted in a significant increase in the rate of gas production, true digestibility, total concentration of VFA, microbial biomass, rate of HCN disappearance, thiocyanate appearance and cyanide percentage conversion into thiocyanate. However, there were no effects of sulfur supplementation at 0.2, 0.5 and 1% to cassava hay. The finding suggests the utilization of cassava foliage for rumen microorganisms in terms of fermentation and HCN detoxification could be improved by sulfur supplementation of 0.5% of DM.

• Korean Journal of Microbiology
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• 제44권2호
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• pp.116-121
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• 2008

In my search of microbial source of novel enzymes, a marine actinomycetes, A1460, producing haloperoxidase was isolated from macroalgae from south sea, Korea and studied for physiological and biochemical properties. The haloperoxidation reaction was followed by the bromination of phenol red in the presence of hydrogen peroxide and potassium bromide. The haloperoxidase was partially purified from the cell extract with $35\sim75%$ ammonium sulfate precipitation, High-Q anion exchange chromatography, gel filtration chromatography, hydroxyapetite chromatography and hydrophobic interaction chromatography to a yield of 42% and purification fold of 70. This enzyme showed relatively high heat stability without losing 50% of activity after 1 hr incubation at $60^{\circ}C$. The highest activity was found at $45^{\circ}C$, and the optimal pH was about pH 7, but higher stability was observed at pH 8. Azide and cyanide ion showed strong inhibition at less than 1 $\mu M$ level suggesting that the enzyme was Fe ion dependent haloperoxidase.

• Molecules and Cells
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• 제23권3호
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• pp.370-378
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• 2007

A superoxide dismutase was purified 62-fold in seven steps to homogeneity from Methylobacillus sp. strain SK1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. The final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was estimated to be 44,000. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,100. The isoelectric point of the purified enzyme was found to be 4.4. Maximum activity of the enzyme was measured at pH 8. The enzyme was stable at pH range from 6 to 8 and at high temperature. The enzyme showed an absorption peak at 280 nm with a shoulder at 292 nm. Hydrogen peroxide and sodium azide, but not sodium cyanide, was found to inhibit the purified enzyme. The enzyme activity in cell-free extracts prepared from cells grown in manganese-rich medium, however, was not inhibited by hydrogen peroxide but inhibited by sodium azide. The activity in cell extracts from cells grown in iron-rich medium was found to be highly sensitive to hydrogen peroxide and sodium azide. One mol of native enzyme was found to contain 1.1 g-atom of iron and 0.7 g-atom of manganese. The N-terminal amino acid sequence of the purified enzyme was Ala-Tyr-Thr-Leu-Pro-Pro-Leu-Asn-Tyr-Ala-Tyr. The superoxide dismutase of Methylobacillus sp. strain SK1 was found to have antigenic sites identical to those of Methylobacillus glycogenes enzyme. The enzyme, however, shared no antigenic sites with Mycobacterium sp. strain JC1, Methylovorus sp. strain SS1, Methylobacterium sp. strain SY1, and Methylosinus trichosproium enzymes.

• Korean Journal of Optics and Photonics
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• 제19권1호
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• pp.54-59
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• 2008

We report on the measurements of absorption spectra from acetylene ($^{12}C_2H_2$) and hydrogen cyanide ($H^{13}C^{14}N$) for wavelength reference in the C-band (conventional band) region using a supercontinuum optical source generated from a mode-locked $Cr^{4+}$:YAG laser. The center wavelength of the mode-locked $Cr^{4+}$:YAG laser was 1510 nm and the pulse duration was 75 fs at 100 MHz repetition rate. The supercontinuum source achieved a flatness of ${\pm}5dB$ over a wavelength range of more than 400 nm, using a 20 m long photonic crystal fiber. The measured absorption spectra from acetylene ($^{12}C_2H_2$) and hydrogen cyanide ($H^{13}C^{14}N$) had more than 50 lines and were analyzed for wavelength standardization.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.100.2-101
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• 2003

The global regulator, GacS (global antibiotic and cyanide sensor kinase), was required for the increased resistance to hydrogen peroxide occurring as cultures of the rhizobacterium, P. chlororaphis O6, matured. Specific stationary-phase peroxidase and catalase isozymes were absent in the GacS mutant, whereas a manganese-superoxide dismutase isozyme was expressed earlier and to a great extent than wild type. In the wild type cell, transcript accumulation of rpoS was higher in late logarithmic-phase cells than cells from mid logarithmic- or stationary-phase. Transcripts from rpoS in the GacS mutant were reduced in each of these growth phases compared to the wild type expression. The down stream sequence from rpoS lacked sequences encoding a small RNA, rsmZ, found in other pseudomonads and implicated in control of genes activated by the GacS system. These findings suggest that GacS-mediated regulation of RpoS plays role in control of oxidative stress in P. chlororaphis O6 by as yet an unknown mechanism.

• Journal of Sensor Science and Technology
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• 제17권2호
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• pp.143-146
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• 2008

Polymer-coated film SAW sensors have been fabricated and their sensing properties for toxic chemicals have been extensively investigated. Four types of the toxic chemical compounds of hydrogen cyanide(AC), carbonyl dichloride(CG), pinacolyl methylfluorophosphonate(GD), 2,2'-dichlorodiethylthio ether(HD) were used as target gases. SAW sensors using five different kinds of polymers were used to detect toxic chemicals and their gas sensing characteristics were investigated. The polymers used as the sensing materials were polyisobutylene(PIB), polyepichlorohydrin(PECH), polydimethylsiloxane(PDMS), polybutadiene(PBD) and polyisoprene(PIP). The recommendable mixing ratio of PIB, PECH, PDMS, PBD and PIP to solvents were 1:30, 1:40, 1:10, 1:30 and 1:30, respectively. The sensing characteristics of the SAW sensors were measured by using E-5061A network analyzer.

• The Plant Pathology Journal
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• 제34권3호
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• pp.241-249
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• 2018

Commercial biocontrol of microbial plant diseases and plant pests, such as nematodes, requires field-effective formulations. The isolate Pseudomonas chlororaphis O6 is a Gram-negative bacterium that controls microbial plant pathogens both directly and indirectly. This bacterium also has nematocidal activity. In this study, we report on the efficacy of a wettable powder-type formulation of P. chlororaphis O6. Culturable bacteria in the formulated product were retained at above $1{\times}10^8$ colony forming units/g after storage of the powder at $25^{\circ}C$ for six months. Foliar application of the diluted formulated product controlled leaf blight and gray mold in tomato. The product also displayed preventative and curative controls for root-knot nematode (Meloidogyne spp.) in tomato. Under laboratory conditions and for commercially grown melon, the control was at levels comparable to that of a standard commercial chemical nematicide. The results indicated that the wettable powder formulation product of P. chlororaphis O6 can be used for control of plant microbial pathogens and root-knot nematodes.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.271-275
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• 1999

The intracellular superoxide dismutase (SOD) from Staphylococcus sciuri was isolated to homogeneity by continuous steps, including ammonium sulfate fractionation, DEAE-ion-exchange chromatography, gel filtration, and phenyl hydrophobic gel chromatography. Pure SOD had a specific activity of 4,625 U/mg and was purified 158-fold with a yield of 31 % from a cell free extract. The molecular weight of the purified SOD was determined to be approximately 35.5 kDa by gel filtration and the enzyme was also shown to be composed of dimeric subunits on denaturing SDS-PAGE. The enzyme activity remained stable at pH 5 to 11 and also to heat treatment of up to $50^{\circ}C$ at pH 7.8, with 80% relative activity. The enzyme was insensitive to cyanide, hydrogen peroxide, and azide, indicating that it is a manganese-containing SOD. The EPR spectrum showed the enzyme containing manganese as a cofactor.

• Research in Plant Disease
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• 제24권2호
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• pp.162-169
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• 2018

The GacS/GacA system in the root colonizer Pseudomonas chlororaphis O6 is a key regulatory system of many traits relevant to the plant probiotic nature of this bacterium. The work in this paper elucidates proteins using proteomics approach in P. chlororaphis O6 under the control of the cytoplasmic regulatory protein, GacA. A gacA mutant of P. chlororaphis O6 showed loss in production of phenazines, acyl homoserine lactones, hydrogen cyanide, and protease, changes that were associated with reduced in vitro antifungal activity against plant fungal pathogens. Production of iron-chelating siderophore was significantly enhanced in the gacA mutant, also paralleling changes in a gacS mutant. However, proteomic analysis revealed proteins (13 downregulated and 7 upregulated proteins in the mutant compared to parental strain) under GacA control that were not apparent by a proteomic study of a gacS mutant. The putative identity of the downregulated proteins suggested that a gacA mutant would have altered transport potentials. Notable would be a predicted loss of type-VI secretion and PEP-dependent transport. Study of mutants of these GacA-regulated proteins will indicate further the features required for probiotic potential in this rhizobacterium.