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Purification of two isofunctional hydrolases (EC 3.7.1.8) in the degradative pathway for dibenzofuran inSphingomonas sp. strain RW1

Abstract

Sphingomonas sp. strain RW1, when grown in salicylate-salts medium, synthesized the enzymes for the degradation of dibenzofuran. The reaction subsequent tometa cleavage of the first benzene ring was found to be catalyzed by two isofunctional hydrolases, H1 and H2, which were purified by chromatography on anion exchange, hydrophobic interaction and gel filtration media. Each enzyme was able to hydrolze 2-hydroxy-6-oxo-6-(2-hydroxyphenyl)hexa-2,4-dienoate and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate to produce salicylate and benzoate, respectively. SDS/PAGE of each purified enzyme showed a single band ofM r 31 000 (H1) or 29 000 (H2). The N-terminal amino acid sequences of the two proteins showed 50% homology.

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Abbreviations

DHB:

2,3-dihydroxybiphenyl

DSM:

German Culture Collection (Braunschweig)

FPLC:

protein liquid chromatograph(y)

HOHPDA:

2-hydroxy-6-oxo-6-(2-hydroxyphenyl)hexa-2,4-dienoate

HOPDA:

2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate

THB:

2,2′,3-trihydroxybiphenyl

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Correspondence to Alasdair M. Cook.

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Bünz, P.V., Falchetto, R. & Cook, A.M. Purification of two isofunctional hydrolases (EC 3.7.1.8) in the degradative pathway for dibenzofuran inSphingomonas sp. strain RW1. Biodegradation 4, 171–178 (1993). https://doi.org/10.1007/BF00695119

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Key words

  • dibenzofuran
  • metabolism of dibenzofuran pathway intermediates
  • 2,2′,3-trihydroxybiphenyl
  • meta cleavage enzymes
  • hydrolysis of a C-C bond (EC 3.7.1.8)
  • HOHPDA hydrolases (EC 3.7.1.8)