Purification of two isofunctional hydrolases (EC 3.7.1.8) in the degradative pathway for dibenzofuran in Sphingomonas sp. strain RW1.
Sphingomonas sp. strain RW1, when grown in salicylate-salts medium, synthesized the enzymes for the degradation of dibenzofuran. The reaction subsequent to meta 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 hydrolyze 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 of M(r) 31,000 (H1) or 29,000 (H2). The N-terminal amino acid sequences of the two proteins showed 50% homology.
Bünz
P V
PV
Institute of Microbiology, ETH-Zentrum, Zürich, Switzerland.
Falchetto
R
R
Cook
A M
AM
D041963:Bacteroidetes / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D001565:Benzoates / Q000378:metabolism; D001572:Benzofurans / Q000378:metabolism*; D019817:Benzoic Acid; D001673:Biodegradation, Environmental; D002396:Catechols / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D006867:Hydrolases / Q000235:genetics / Q000302:isolation & purification*; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000378:metabolism; D012459:Salicylates / Q000378:metabolism; D020156:Salicylic Acid
D001565:Benzoates; D001572:Benzofurans; D002396:Catechols; D005231:Fatty Acids, Unsaturated; D007527:Isoenzymes; D012459:Salicylates; C085250:2-hydroxy-6-oxo-6-(2-hydroxyphenyl)hexa-2,4-dienoate; C018971:2-hydroxy-6-oxo-6-phenyl-2,4-hexadienoic acid; D019817:Benzoic Acid; C023614:dibenzofuran; C067814:3-(2-hydroxyphenyl)catechol; D010105:Oxygenases; D006867:Hydrolases; C062907:2,6-dioxo-6-phenylhexa-3-enoate hydrolase; D020156:Salicylic Acid