Isolation and characterization of an endo-beta-D-glucuronidase from the fungus Kobayasia nipponica.
An endo-beta-D-glucuronidase was isolated and characterized from Kobayasia nipponica. The enzyme was purified by ammonium sulfate fractionation, CM-Sephadex chromatography, gel filtration with Sephacryl S-200, and heparin-Sepharose chromatography. The enzyme shows the following properties: optimum pH 5.0, thermal stability below 37 degrees C, pH stability 5-6, optimum temperature 45-55 degrees C, and Km 0.12% for L-idurono-D-glucuronan (protuberic acid (PA), L-IdUA:D-GlcUA = 1:2) from Kobayasia nipponica, 0.19% for PF (L-IdUA:D-GlcUA = 1:3) from Pseudocolus fusiformis, and 0.23% for (1-4)-beta-D-glucuronan(mucoric acid) from Mucor mucedo as determined from Hofstee plots. The molecular weight values estimated by gel filtration through Sephacryl S-200 and Sephadex G-50 and by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate were 10,500 and 10,200, respectively. The endo-beta-D-glucuronidase was inactive towards several glycosaminoglycans.
D001487:Basidiomycota / Q000201:enzymology*; D002850:Chromatography, Gel; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D005966:Glucuronidase / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D013696:Temperature
D008345:Manganese; D005966:Glucuronidase