• Title/Summary/Keyword: Phenol Red

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Purification and Characterization of the Laccase Involved in Dye Decolorization by the White-Rot Fungus Marasmius scorodonius

  • Jeon, Sung-Jong;Lim, Su-Jin
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1120-1127
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    • 2017
  • Marasmius scorodonius secretes an extracellular laccase in potato dextrose broth, and this enzyme was purified up to 206-fold using $(NH_4)_2SO_4$ precipitation and a Hi-trap Q Sepharose column. The molecular mass of the purified laccase was estimated to be ~67 kDa by SDS-PAGE. The UV/vis spectrum of the enzyme was nontypical for laccases, and metal content analysis revealed that the enzyme contains 1 mole of Fe and Zn and 2 moles of Cu per mole of protein. The optimal pH for the enzymatic activity was 3.4, 4.0, and 4.6 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonate) (ABTS), guaiacol, and 2,6-dimethoxy phenol as the substrate, respectively. The optimal temperature of the enzyme was $75^{\circ}C$ with ABTS as the substrate. The enzyme was stable in the presence of some metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Ni^{2+}$, $Mg^{2+}$, $Mn^{2+}$, $Ba^{2+}$, $Co^{2+}$, and $Zn^{2+}$ at a low concentration (1 mM), whereas $Fe^{2+}$ completely inhibited the enzymatic activity. The enzymatic reaction was strongly inhibited by metal chelators and thiol compounds except for EDTA. This enzyme directly decolorized Congo red, Malachite green, Crystal violet, and Methylene green dyes at various decolorization rates of 63-90%. In the presence of 1-hydroxybenzotriazole as a redox mediator, the decolorization of Reactive orange 16 and Remazol brilliant blue R was also achieved.

Kinetic Properties of Manganese Peroxidase from the Mushroom Stereum ostrea and its Ability to Decolorize Dyes

  • Praveen, K.;Usha, K.Y.;Viswanath, Buddolla;Reddy, B. Rajasekhar
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1540-1548
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    • 2012
  • Manganese peroxidase (MnP) was isolated from the culture filtrate of the wood log mushroom Stereum ostrea (S. ostrea), grown on Koroljova medium, and then purified by ammonium sulfate [70% (w/v)] fractionation, DEAE-cellulose anion exchange chromatography, and Sephadex G-100 column chromatography, with an attainment of 88.6-fold purification and the recovery of 22.8% of initial activity. According to SDS-PAGE the molecular mass of the MnP was 40 kDa. The optimal pH and temperature were found to be 4.5 and $35^{\circ}C$, respectively. The enzyme was stable even after exposure to a pH range of 4.5 to 6.0, and at temperatures of up to $35^{\circ}C$ at a pH of 4.5 for 1h. The $K_m$ and $V_{max}$ values for the substrate phenol red were found to be $8{\mu}m$ and 111.14 U/mg of protein, respectively. The MnP also oxidized other substrates such as guaiacol, DMP, and veratryl alcohol. Sodium azide, EDTA, SDS, $Cu^{2+}$, and $Fe^{2+}$, at 1-5 mM, strongly inhibited enzyme activity, whereas $Ca^{2+}$ and $Zn^{2+}$ increased enzyme activity. The participation of the purified enzyme in the decolorization of dyes suggests that S. ostrea manganese peroxidase could be effectively employed in textile industries.

Antimicrobial susceptibility features of porcine respiratory bacterial pathogens by modified broth dilution method (돼지 호흡기질병 병인체의 항균제 감수성 조사)

  • 송동준;서동균;이춘식;배영찬;김원일;김봉환
    • Korean Journal of Veterinary Service
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    • v.23 no.1
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    • pp.19-28
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    • 2000
  • There are several main antibacterial susceptibility tests, such as agar dilution method, broth dilution method and disk diffusion technique. Especially, for minimal inhibitory concentration (MIC) test, agar dilution method has been widely used. But that method is so complicated and bothering that it's difficult to treat a large amount of strains. On the other hand, modified broth dilution method(add 1% glucose and 0.018% phenol red as a pH indicator to broth) is fast and easy to perform. Most of all, it can visualize the result by color. The MICs of 22 antibiotics Including penicillins, aminoglycosides, cephalothin, chloramphenicol, lincomycin, ceftiofur, vancomycin and quinolones, erythromycin, colistin. sul-fadimethoxine, trimethoprim for arcanobacterium pyogenes 14 strains, actinobacillus pleuropneu-moniae 41 strains and pasteurella multocida 37 strains, which were collected from porcine during 1996 ∼ 1999, were determined by modified broth dilution method. Actinobacillus pleuropneumoniae was highly susceptible to all kinds of quinolones such as ciprofloxacin, enrofloxacin and norfloxacin and to all aminoglycosides, like gentamicin, apramycin, kanamycin and ampicillin, cephalothin and ceftiofur. But It was quite resistant to solfadimethoxin, colistin and vancomycin. Pasteurella multocida was found to have high susceptibility to ampicillin, cephalothin, chlorampenicol and gentamicin but had mid-degree susceptibility to other aminoglycosides. In addition, it was susceptible to norfloxacin and nalidixic acid, but not to newer fluoroquinolone like ciprofloxacin and enrofloxacin and it was resistant to colistin and kanamycin. Arcanobacterium pyogenes was highly susceptible to most of quinolones such as cipoofloxacin, enrofloxacin and norfloxacin and gentamicin and penicillin G. But it also obtained high resistance against the early quinolone, nalidixic acid and aminoglycosides such as amikacin, apramycin and kanamycin and erythromycin, chlorampenicol, tetracyclin and vancomycin.

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Effect of Adhesives and Finger Pitches on Bending Creep Performances of Finger-Jointed Woods

  • Park, Han-Min;Oh, Seong-Won;Byeon, Hee-Seop
    • Journal of the Korean Wood Science and Technology
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    • v.33 no.5 s.133
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    • pp.57-65
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    • 2005
  • Following our previous reports for finger-jointed woods with various finger profiles studied for the efficient use of small diameter logs and woods containing various defects, twelve types of finger-jointed woods glued with three kinds of adhesives and with two sizes of finger pitches were made with sitka spruce and red pine. The effects of the adhesives and finger pitches on bending creep performances of finger-jointed woods were investigated. The shape of creep curves differed among the used adhesives and finger pitches of finger-jointed woods for both tested species. Their creep curves showed a linear behavior beyond about one hour, and the N values fitted to power law increased with increasing finger pitches. The initial deformation increased with increasing finger pitches, regardless of the tested species and kinds of adhesives, whereas the effect of finger pitches on the creep deformation was not clear. For finger-jointed woods glued with polyvinyl acetate (PVAc) resin, creep failure occurred in 106 hours after the load was applied. And the difference of the creep compliance between finger-jointed woods glued with resorcinol-phenol formaldehyde (RPF) resin and aqueous vinyl urethane (AVU) resin was small. The ratios for creep performances of finger-jointed woods glued with RPF resin and AVU resin versus solid wood were higher in creep deformation than initial deformation for both species, and the difference between both adhesives was not found. The relative creep decreased with increasing finger pitches, and the marked differences was not found between RPF resin and AVU resin.

A Study on the General Characteristics and Instrumental Analysis of Natural Omija Extract

  • Sung, Ki-Chun;Kim, Ki-Jun;Kim, Yong-Ryul;Nam, Sang-Sung
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.2
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    • pp.225-232
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    • 2013
  • Omija component was known to possess natural odor, taste, color, and various general characteristics. Omija extraction was extracted using ethanol as a solvent. Omija extract showed a red-purple color of some viscous liquid state. Some conclusions from natural Omija extract were obtained as follow. The result of antimicrobial experiment to add Omija extract, the number of microbial population showed negative reaction from 3 days after it cultivated. This phenomenon could confirm that Omija component affected to antimicrobial effect. The result of dyeing experiment to add Omija extract, fiber dyeing effect showed with some ivory color after dyed to cotton and silk. Also, this phenomenon could confirm that Omija component affected to natural dyeing effect from observated dye state with biological microscope(BM). The result of instrumental analysis, inorganic components of K(109.60ppm), Na(3.500ppm), Ca(1.205ppm), Mg(0.900ppm), Li(0.350ppm), Si(0.380ppm), Cu(0.250ppm), Fe(0.125ppm), Zn(0.090ppm), etcs from Omija were ascertained with ICP/OES, and organic components of benzene(10.808), borny lacetate(11.289), phenol(14.183), ${\beta}$-terpinene(15.840), ${\alpha}$-terpinolene(17.616) etcs from Omija were ascertained with GC/MSD.

Relief Effect of Medicinal Herb Mixture HAE Series on Respiratory Symptoms (호흡기 증상에 대한 복합제 HAE의 완화 효과)

  • Chang, Bo Yoon;Kim, Da Eun;Han, Ji Hye;Lee, Joo Seok;Kim, Dae Sung;Cho, Hyoung Kwon;Kim, Sung Yeon
    • Korean Journal of Pharmacognosy
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    • v.46 no.4
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    • pp.334-341
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    • 2015
  • HAE series (HAE 01 - 06), are the mixtures of medicinal plants; Lonicerae Folium et Caulis (Lonicera japonica), Scutellariae Radix (Scutellaria baicalensis), Adenophorae Radix (Adenophora triphylla var. japonica), Polygonati Oddorati Rhizoma (Polygonatum odoratum var. pluriflorum), Platycodi Radix (Platycodon grandiflorum), Liriopis seu Ophiopogonis Tuber (Liriope platyphylla). HAE 06 showed the significant antitussive effect in HAE series. HAE 04, 05, and 06 increased the effect of expectorant measured by secretion of phenol red in rats. All tested HAE series were observed to have the anti-oxidant activity determined by the scavenging activity. HAE 01, 03 and 06 were significantly reduced the production of nitric oxide (NO) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) increased by LPS. HAE 06 was selected based on the results and done further experiments. HAE 06 inhibited the number of coughing induced by citric acid in dose dependent manner. HAE 06 effectively increased the effect of expectorant. HAE 06 exhibited anti-inflammatory effects by inhibiting the production of NO and TNF-${\alpha}$. In addition, HAE 06 significantly suppressed inducible Cyclooxygenase-2 (COX-2). Especially HAE 06 has the highest effect in alleviating the respiratory symptoms. Moreover, the enhancement antioxidant activity and reduction inflammation partly were attributed to it. HAE 06 may become the promising candidate for the treatment of respiratory symptom in the future.

Screening and Partial Purification of Haloperoxidase from Marine Actinomycetes (해양방선균으로부터 Haloperoxidase의 검색과 특성)

  • Cho, Ki-Woong
    • Korean Journal of Microbiology
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    • v.44 no.2
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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.

COMPARISON OF THE SEALING ABILITY OF VARIOUS RETROGRADE FILLING MAIERIALS (수종의 역충전 재료의 치근단 밀폐력 비교)

  • 황윤찬;강인철;황인남;오원만
    • Restorative Dentistry and Endodontics
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    • v.26 no.5
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    • pp.379-386
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    • 2001
  • This study was performed to evaluate the sealing ability of various retrograde filling materials by using bacterial penetration and dye penetration test. One hundred and forty extracted human teeth with single, straight canals and mature apiece were collected and used for this study. All canals were instrumented using an engine driven Ni-Ti file (ProFile). After removing 3mm from the apex of tooth, a standardized 3mm root end cavity was prepared using an ultrasonic instrument. The 70 teeth were randomly divided into 7 groups : 6 groups for retrograde filling using Super-EBA, ZOE, Chelon-Silver, IRM, ZPC and amalgam. The 7th group was used as a negative control. Nail varnish was applied to all external root surfaces to the level of the reseated root ends to prevent lateral microleakages. The specimens were then sterilized in an ethylene oxide sterilizer for 24 hours. 2 mm of the reseated root was immersed in a culture chamber containing a Tripticase Soy Broth with a phenol red indicator. The coronal access of each specimen was inoculated every 72 hours with suspension of Proteus vulgaris. The culture media were observed every 24hours for color change indicating bacterial contamination. The specimens were observed for 4weeks. The remaining 70 teeth were submitted to a dye penetration test. The canals of all teeth were first sealed with AH26 and obturated using an Obtura II system. Root resection, root end preparation and retrograde filling was performed as above. All specimens were suspended in 2% methylene blue dye for 72 hours before being ion gitudinally split. The degree of dye penetration was then measured using a stereomicroscope at 10 magnification and evaluated. The results were as floows : 1. In the bacterial penetration, the degree of leakage was the lowest in the Super-EBA, followed by, in ascending order, ZOE, Chelon-Silver IRM and ZPC. The amalgam showed highest bacterial leakage of all(p<0.01). 2. In the dye penetration, the degree of microleakage was the lowest in the Chelon-Silver and Super-EBA, followed by, in ascending order, IRM, ZPC. The ZOE and amalgam showed the highest microleakage of all (p<0.05). These results suggested that the eugenol based cement, Super-EBA, have excellent sealing ability as a retrograde filling material.

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The effect of progesterone and 17-β estradiol on membrane-bound HLA-G in adipose derived stem cells

  • Moslehi, Akram;Hashemi-beni, Batool;Moslehi, Azam;Akbari, Maryam Ali;Adib, Minoo
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.4
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    • pp.341-346
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    • 2016
  • Membrane-bound HLA-G (mHLA-G) discovery on adipose derived stem cells (ADSCs) as a tolerogenic and immunosuppressive molecule was very important. Many documents have shown that HLA-G expression can be controlled via some hormones such as progesterone (P4) and estradiol (E2). Therefore, this study was designed to evaluate progesterone and estradiol effects on mHLA-G in ADSCs at restricted and combination concentrations. Three independent cell lines were cultured in complete free phenol red DMEM and subcultured to achieve sufficient cells. These cells were treated with P4, E2 and P4 plus E2 at physiologic and pregnancy concentrations for 3 days in cell culture conditions. The HLA-G positive ADSCs was measured via monoclonal anti HLA-G-FITC/ MEMG-09 by means of flow cytometry in nine groups. Data were analyzed by one way ANOVA and Tukey's post hoc tests. There were no significant values of the mean percentage of HLA-G positive cells in E2-treated and the combination of P4 plus $E_2-treated$ ADSCs compared to control cells (p value>0.05) but P4 had a significant increase on mHLA-G in ADSCs (p value<0.05). High P4 concentration increased mHLA-G but E2 and the combination of P4 plus E2 could not change mHLA-G on ADSCs.

The effect of $Ni^{2+}$ on the intracellular $Ca^{2+}$ increase of the mouse early 2-cell embryos (생쥐 초기 2-세포 배에서 세포 내 칼슘 농도의 변화에 $Ni^{2+}$이 미치는 영향)

  • Yoon, Sook-Young;Lee, Eun-Mi;Bae, In-Ha
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.4
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    • pp.269-280
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    • 2003
  • Objective: We reported the overcoming effect of $Ni^{2+}$ on the in vitro 2-cell block of mouse embryos. In this study, we aim to investigate whether $Ni^{2+}$ should induce intracellular $Ca^{2+}$ transient in the mouse embryos. Materials and Methods: Embryos were collected at post hCG 32hr from the oviduct of the ICR mouse and cultured in M2 medium omitted phenol red. Intracellular $Ca^{2+}$ was checked by using a confocal laser scanning microscope and fluo-3AM by using various intracellular $Ca^{2+}$ antagonists. Results: In 1mM $Ni^{2+}$ treated medium which contained $Ca^{2+}$(1.71mM), 75.7% of the embryos showed $[Ca^{2+}]i$ transient about 200 sec later. In the $Ca^{2+}$-free medium, 69.8% of the embryos showed $[Ca^{2+}]i$ transient. In U73122, phospholipaseC(PLC) inhibitor (5uM, 10min) pretreated group, 33.3% of the embryos showed $[Ca^{2+}]i$ transient. Heparine, inositol 1, 4, 5-triphosphate receptor(IP3R) antagonist preinjected embryos showed no response with 1mM $Ni^{2+}$. In danthrolene treatment, ryanodine receptor(RyR)-antagonist, 43% embryos showed $[Ca^{2+}]i$ transient but they showed delayed response about 340sec in the presence of $Ca^{2+}$. Conclusions: Summing up the above results, $Ni^{2+}$ seems to induce $Ca^{2+}$-release from the $Ca^{2+}$-store even in the $Ca^{2+}$-free medium. IP3 receptors of the mouse 2-cell embryos might have an essential role for the intracellular $Ca^{2+}$ increase by $Ni^{2+}$.