title: β-Glucuronidase-coupled assays of glucuronoyl esterases.
abstract: Glucuronoyl esterases (GEs) are microbial enzymes with potential to cleave the ester bonds between lignin alcohols and xylan-bound 4-O-methyl-d-glucuronic acid in plant cell walls. This activity renders GEs attractive research targets for biotechnological applications. One of the factors impeding the progress in GE research is the lack of suitable substrates. In this work, we report a facile preparation of methyl esters of chromogenic 4-nitrophenyl and 5-bromo-4-chloro-3-indolyl β-D-glucuronides for qualitative and quantitative GE assay coupled with β-glucuronidase as the auxiliary enzyme. The indolyl derivative affording a blue indigo-type product is suitable for rapid and sensitive assay of GE in commercial preparations as well as for high throughput screening of microorganisms and genomic and metagenomic libraries.
authors.affiliation: Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-845 38 Bratislava, Slovak Republic. Electronic address: lucia.franova@savba.sk.
authors.forename: Lucia
authors.initials: L
authors.lastname: Fraňová
authors.affiliation: Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-845 38 Bratislava, Slovak Republic. Electronic address: vladimir.puchart@savba.sk.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Puchart
authors.affiliation: Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-845 38 Bratislava, Slovak Republic. Electronic address: peter.biely@savba.sk.
authors.forename: Peter
authors.initials: P
authors.lastname: Biely
mesh_terms: D001419:Bacteria / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry*; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry*; D002473:Cell Wall / Q000737:chemistry; D005965:Glucuronates / Q000737:chemistry*; D005966:Glucuronidase / Q000737:chemistry*; D010944:Plants / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D005965:Glucuronates; D002265:Carboxylic Ester Hydrolases; D005966:Glucuronidase
keywords: CE15; Chromogenic substrate; Coupled enzyme reaction; Glucuronoyl esterase; β-Glucuronidase