• Title/Summary/Keyword: methanol dehydrogenase

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A Methylobacillus Isolate Growing Only on Methanol (메탄올만 이용하여 성장하는 Methylobacillus의 분리 및 특성)

  • 김시욱;김병홍;김영민
    • Korean Journal of Microbiology
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    • v.29 no.4
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    • pp.250-257
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    • 1991
  • An obligate methanol-oxidizing bacterium, Methylobacillus sp. strain SK1, which grows only on methanol was isolated from soil. The isolate was nonmotile Gram-negtive rod. It does not have internal membrane system. The colonies were small, whitish-yellow, and smooth. The guanine plus cytosine content of the DNA was 48 mol%. Cellular fatty acids consisted predominantly of large amounts of straight-chain saturated $C_{16:0}$ acid and unsaturated $C_{16:1}$ acid. The major ubiquinone was Q-8, and Q-10 was present as minor component. The cell was obligately aerobic and exhibited catalase, but no oxidase, activity. Poly-.betha.-hydroxybutyrate, endospores, or cysts were not observed. the isolate could grow only on methanol in mineral medium. Growth factors were not required. The isolate was unable to use methane, formaldehyde, formate, methylamine, and several other organic compounds tested as a sole source of carbon and energy. Growth was optimal at 35.deg.C and pH 7.5. It could not grow at 42.deg.C. The doubling time was 1.2h at 30.deg.C when grown with 1.0%(v/v) methanol. The growth was not affected by antibiotics inhibiting cell wall synthesis and carbon monoxide but was completely suppressed by those inhibiting protein synthesis. Methanol was found to be assimilated through the ribulose monophosphate pathway. Cytochromes of b-, c-, and o- types were found. Cell-free extracts contained a phenazine methosulfate-linked methanol dehydrogenase activity, which required ammonium ions as an activator. Cells harvested after the late exponential phase seemed to contain blue protein.ein.

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Activities of Antioxidation and Alcohol Dehydrogenase Inhibition of Methanol Extracts from Some Medicinal Herbs (약용식물 추출물에 대한 항산화성과 알코올 탈수소효소 저해성 연구)

  • 문지숙;김선재;박윤미;황인식;김의형;박정욱;박인배;김상욱;강성국
    • Food Science and Preservation
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    • v.11 no.2
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    • pp.201-206
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    • 2004
  • The activities of antioxidation and alcohol dehydrogenase in hibitionin methanol extracts of thirty two medical herbs were tested using the method of DPPH activity, nitrite scavenging effect and alcohol dehydrogenase assay in vitro. In DPPH method, Eugenia caryophyllata, Thea sinensis, Paeonia suffruticosa, Alnus japonica showed over 90 % of free radical scavenging activities. The nitrite scavenging ability appeared Zanthoxylum bungeanum, Alnus japonica, Thea sinensis, Hovenia dulcis(cortex) and Illicium verum showed the high value. In connection with in vivo alcohol metabolism, thirteen medicinal herbs were screened for inhibition. As a reasult, we found significant inhibition of ADH by methanolic extracts of Glycyrrhiza uralensis, Pueraria thunbergiana(radix), Alnus japonica. These results indicate that the antioxidative effect was strongly related with alcohol dehydrogenase inhibitor; Thea sinensis and Alnus japonica.

Effect of Extraction Solvent on the Physiological Properties of Korean Pear Peel (Pyrus pyrifolia cv. Niitaka) (추출 용매를 달리한 한국산 배(Pyrus pyrifolia cv. Niitaka) 과피의 생리 활성)

  • Park, Ji-Su;Han, Inhwa
    • Korean Journal of Food Science and Technology
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    • v.47 no.2
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    • pp.254-260
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    • 2015
  • The effect of the extraction solvent on the physiological properties of the peel of the Korean pear (Pyrus pyrifolia cv. Niitaka) was evaluated. The total phenol content was highest in the 80%(wt) methanol extract, whereas flavonoid content was highest in the 80% ethanol extract. The antioxidant activities of the extracts were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging abilities, and their reducing power. The water and 80% methanol extracts of the pear peel had highest DPPH and ABTS radical scavenging activities, and reducing power, respectively. The inhibition of ${\alpha}$-glucosidase was highest in the 80% methanol extract, and alcohol dehydrogenase activity was highest in the water extract. All three extracts had similar antimicrobial activity. Because water, 80% ethanol, and 80% methanol extracts exhibited high activities in different assays of physiological properties, each solvent could be used for specific purposes.

Purification and Characterization of Alcohol Dehydrogenase from Acetobacter sp. KM (Acetobater sp.KM Alcohol Dehydrogenase의 분리 및 특성)

  • 전홍성;차영주
    • KSBB Journal
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    • v.10 no.1
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    • pp.30-37
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    • 1995
  • Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. KM. The enzyme was solubilized and extracted with Triton X-100 and purified using the Mono-Q ion exchange chromatography and Superose 12 gel filtration chromatography. The enzyme was purified to 12-fold with a yield of 30%. The molecular weight of the purified enzyme was to be 335 KDa. SDS-PAGE of the enzyme showed two subunits with molecular weights of 79 KDa and 49 KDa. It indicated that the enzyme consisted of three subunits of the 79 KDa and two subunits of the 49 KDa. The purified .ADH preferentially oxidized straight chain aliphatic alcohol except methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidized. The apparent Km for ethanol was 1.04 mM and the optimum pH and temperature were 5.0∼6.0 and 32$^{\circ}C$, respectively. V2O5 and divalent cation such as ZnCl2 and NiCl2 inhibited enzymatic activity.

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Effects of Lycii Fructus on Primary Cultured Chicken Brain Cells

  • Park, Mi-Jung;Chu, Eun-Hye;Lee, Heun-Pa;Kim, Young-Choong
    • Archives of Pharmacal Research
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    • v.14 no.4
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    • pp.325-329
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    • 1991
  • Effects of Lycii Fructus on primary cultured chicken embryonic brain cells were studied by microscopic observation, determination of the activity of pyruvate dehydrogenase complex (PDHC), and syntheses of protein, RNA and DNA. The brain cells were prepared from the brains or 10-day-old chicken embryos and cultured with a deficient medium. The activity of PDHC in the brain cells cultured with a deficient medium was increased to 1.8 times by the addition of $30\;{\mu}g/ml$ of the total methanol extract of Lycii Fructus. To seek the active fraction, total methanol extract was further fractionated by the polarity. The survival rate of neuronal cells was significantly increased by the addition of $100\;{\mu}g/ml$ of the buthanol or aqueous fraction. At this concentration, the significant increase of the syntheses of protein and RNA, but not of DNA, indicates that the fractions may act on the neuronal cells which are known to be non-dividing cells.

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Evaluation of Cell Cytotoxicity on the extractives of Magnoliaceae (목련과 수목의 추출성분에 대한 세포독성평가)

  • 김영균;고영남;김용만;양현옥
    • Journal of Korea Foresty Energy
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    • v.20 no.2
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    • pp.1-8
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    • 2001
  • The 90% methanol extracts of eight magnoliaceae plants were collected and tested the cytotoxicity against SK-OV-3 and SiHa cells. Also six pure compounds such as magnonol, honokiol, dihydroxybiphenyl ether, linodenine, anonaine, asimilobine which were previously isolated from Magnolia obovata Thunb. were evaluated the cytotoxicities and their mechanism study using the Lactate dehydrogenase assay(LDH) and FACScan analysis system. Of the tested six compounds, magnonol, honokiol, dihydroxybiphenyl ether showed high cytotoxicities against human cancer cell lines, SK-OV-3 and SiHa cells. In addition, one of the plausible mechanisms of their antitumor activities suggested that they could induce the early stage of apoptosis. For the quantitative analysis, the methanol extractives were fractionated with chloroform, ethylacetate, $H_2O$ and then the ethylacetate fraction was chromatographed on silica gel using n-Hexane ; Acetone(4:1, v/v) as eluent. This fraction was subjected for the quantitative analysis in the HPLC system. The result suggested that the methanol extractives of Magnolia obovata Thunb. contained with magnonol, honokiol, dihydroxybiphenyl ether, 0.9%, 0.3% and 0.24%, respectively.

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Production of Methanol by Resting Cells of Methylosinus trichosporium OB3b (Methylosinus trichosporium OB3b를 이용한 메탄올의 생산)

  • 박성훈;추석열
    • KSBB Journal
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    • v.8 no.4
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    • pp.341-350
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    • 1993
  • As an effort to develop an alternative transportation fuel, the production of methanol from methane gas was studied using the resting cells of an obligatory methanotroph, Methylosinus trichosporium OB3b. The reaction was carried out in high concentration phosphate buffer solutions with the flask-grown cells containing the exclusively cytoplasmic methane monooxygenase (sMMO) activity. The methanol accumulation rate was observed to be 79nmo1/mg·min during the initial 4.5hr. Phosphate-dependent inhibition was found for both sMMO and methanol dehydrogenase (MDH) activities, and the inhibition constants were 185mM and 42mM, respectively. The inhibition mode was noncompetitive. Methanol was found to be very inhibitory to the sMMO activity and the inhibition constant (noncompetitive) was 21mM when propylene was used as substrate. The sMO activity in the resting cells was declined very fast and the rate became very high during the methanol production. These results indicate that the use of M. trichosporium OB3b as a biocatalyst for the methanol production is heavily dependent on the stable maintenance of the whole-cell SMO activity as well as the effective alleviation of product inhibition.

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Purification and Properties of a Membrane-bound Alcohol Dehydrogenase from Acetobacter sp. HA (Acetobacter sp. HA로부터 Membrane-bound Alcohol Dehydrogenase의 정제 및 특성)

  • Yoo, Jin-Cheol;Sim, Jung-Bo;Kim, Heung-Keun;Chun, Hong-Sung;Kim, Sung-Jin
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.78-83
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    • 1994
  • Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneous state fron an acetic acid producing bacteria, Acetobacter sp. HA. The enzyme was purified about 153-fold with an overall yield of 35% from the crude cell extract by solubilization and extraction of the enzyme with Triton X-100 and subsequent fractions by column chromatography. Upon sodium dodecyl sulphate-PAGE, the enzyme showed the presence of three subunits with a molecular mass of 79,000 daltons, 49,000, and 45,000 daltons, respectively. Absorption oxidized aliphatic alcohols with a straight carbon chain except for methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidizable substrates. The apparent $K_m$ for ethanol was 1.38mM. The optimun pH and temperature were 5.0~6.0 and 32${\circ}C$, respectively. $V_2O_5$ and heavy metals such as $ZnCl_2\;and\; NiCl_2$ were inhibitory to the enzyme activity.

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Biotechnology for the Production of Threonine Production (Threonine의 생물공학적 생산)

  • Kim, Kyoung-Ja
    • YAKHAK HOEJI
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    • v.34 no.6
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    • pp.447-456
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    • 1990
  • Various methods are available for the production of L-threonine. The microbial production of L-threonine has been achieved by breeding L-threonine analog-resistant auxotrophic mutants of various bacteria. The enzymatic production of L-threonine has been demonstrated by use of threonine metabolic enzymes such as threonine deaminase, threonine aldolase, or threonine dehydrogenase complex. Threonine synthesis from glycine and ethanol seems to be catalyzed by the enzymes Methanol dehydrogenase(MDH) and Serine hydroxymethyltransferase(SHMT), which was also found to catalyze the aldol condensation of glycine with acetaldehyde. The improved production of L-threonine has been achieved by amplifying the genes for the L-threonine biosynthetic enzymes using recombinant DNA techniques.

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Biochemical Studies on the Chemical Components of Korean Ginseng(III) Effects of Ginseng Components on the Activity of Succinate Dehydrogenase (한국 인삼 성분들에 관한 생화학적 연구(III) 인삼 성분들이 숙신산 탈수소효소의 활동성에 미치는 영향)

  • 장세희;박인원
    • Journal of Ginseng Research
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    • v.1 no.1
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    • pp.29-32
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    • 1976
  • Succinate dehydrogenase was activated by ethyl acetate extract from the methanol extract of white ginseng previously treated with petroleum ether and ethyl ether to remove all highly nonpolar components, and the residual aqueous solution from the ethyl acetate extraction. Also, all of the extracted fractions of red ginseng except the ether extract gave positive results. On the contrary to some suggestions by other workers that alkaloids of ginseng may enhance the succinate degydrogenase activity, our results show that the alkaloids may have exhibited some inhibitory eject on this enzyme.

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