Purification and Characterization of an Ether Bond-Cleaving Enzyme Involved in the Metabolism of Polyethylene Glycol

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概要

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• An ether bond-cleaving enzyme was purified as diglycolic acid (DGA) dehydrogenase associated with 3-(4,5-dimethyl-2-thioazolyl)-2,5-diphenyl-2H tetrazolium bromide and phenazine methosulfate. DGA dehydrogenase was inductively formed in a polyethylene glycol (PEG)-utilizing symbiotic mixed culture, E-1,composed of Sphingomonas terrae and Rhizobium sp. which were grown on PEG 6,000. DGA dehydrogenase was solubilized with 0.5% n-dodecyl-β-D-maltoside from membrane fractions of the mixed culture E-1 and purified almost to homogeneity, as determined by SDS-polyacrylamide gel electrophoresis, by DEAE-Toyopearl column chromatography and Sepharose 6B gel filtration. The molecular weight of the enzyme was determined to be about 40,000-41,000 Da by SDS-polyacrylamide gel electrophoresis and 480,000 by gel filtration on Sepharose 6B in the presence of 0.5% n-dodecyl-β-D-maltoside. The absorption spectrum of the purified enzyme showed two peaks at 345 nm and 430-450 nm in addition to a major peak at 280 nm. The optimal pH and temeperature were 8.9 and 40℃, respectively. The enzyme was stable in the pH range of 7.8 to 9.0 and at below 30℃. The enzyme was activated by neither metal ions nor ammonium ion, and strongly inhibited by 1,4-benzoquinone. The enzyme catalyzed the degradation of DGA, PEG dicarboxylic acids, glycolic acid and glyoxylic acid, but not primary alcohols, ethylene glycol (EG), EG oligomers, PEG 6,000 and aldehydes.

• 社団法人日本生物工学会の論文
• 1997-06-25

著者

• KAWAI Fusako

  Department of Biology, Kobe University of Commerce

• Enokibara Shogo

  Department Of Biology Kobe University Of Commerce

• Kawai Fusako

  Department Of Agricultural Chemistry College Of Agriculture Kyoto University

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• Identification of Reaction Products of Polyethylene Glycol Dehydrogenase
• Aerobic Degradation of Polypropylene Glycol by Corynebacterium sp.
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• Bacterial Assimilation of Polytetramethylene Glycol
• Identification of Polyethylene Glycols (PEGs)-assimilable Bacteria and Culture Characteristics of PEG 6000 Degradation by a Mixed Culture : Micro Degradation of Synthetic Polymers (III) See Reference 1).
• Purification and Characterization of an Ether Bond-Cleaving Enzyme Involved in the Metabolism of Polyethylene Glycol
• Symbiotic Degradation of Polyethylene Glycol (PEG) 20,000-Phthalate Polyester by Phthalate Ester-and PEG 20,000-Utilizing Bacteria
• Production of a Novel Extracellular Polysaccharide by a Bacillus Strain Isolated from Soil
• Inducible or Constitutive Polyethylene Glycol Dehydrogenase Involved in the Aerobic Metabolism of Polyethylene Glycol
• Taxonomical Position of Newly Isolated Polyethylene Glycol-Utilizing Bacteria
• Purification and Characterization of Constitutive Polyethylene Glycol (PEG) Dehydrogenase of a PEC 4000-Utilizing Flavobacterium sp. No. 203
• Identification of the Prosthetic Group and Further Characterization of a Novel Enzyme, Polyethylene Glycol Dehydrogenase

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