A simple, reliable and accurate method for the measurement of alpha-D-glucuronidase in various enzyme preparations. Containing and aldouronic acid substrate and an α-D-glucuronidase control.
INTRODUCTION:
After cellulose, xylan is the second most abundant polysaccharide
found in nature. Xylan is a hemicellulose comprising a β-(1,4)-
linked xylan back bone substituted with various side chains such as
acetyl groups, α-(1-2/3)-linked L-arabinofuranose and α-(1-2)-linked
4-O-methylglucuronic acid. A repertoire of enzymes are required to
effect the complete degradation of plant cell wall polysaccharides in
the utilisation of plant biomass. Hence, complete degradation of xylan
requires the concerted action of various enzymes including endo-β-
(1,4)-xylanases, β-(1,4)-xylosidases, acetylxylan esterases,
α-L-arabinofuranosidases and α-glucuronidases.
The importance of carbohydrate degrading enzymes has increased
over recent years due to the growing focus of obtaining renewable
energy from plant biomass in the form of biofuels such as cellulosic
and hemicellulosic ethanol. Various research programmes are striving
to develop optimised enzyme mixtures or modified microorganisms
to increase the efficiency of biomass conversion and, as such, it is
important that the various enzyme activities can be easily measured.
Simple methods of measuring enzyme activity for some of the well
studied carbohydrate degrading enzymes such as endo-β-(1,4)-
cellulases, endo-β-(1,4)-xylanases, β-(1,4)-xylosidases exist. However
for some enzymes such as α-glucuronidases, current available methods
are cumbersome.
This kit (K-AGLUA) provides a simple method suitable for the specific
measurement of α-D-glucuronidase activity (α-D-glucosiduronate
glucuronohydrolase; EC 3.2.1.139 and xylan α-1,2-glucuronosidase;
EC 3.2.1.131).
KITS:
Kits suitable for performing 50 assays in manual format (or 200 assays
in microplate format) are available from Megazyme. The kits contain
the full assay method plus:
Bottle 1: Aldouronic acid mixture (tri:tetra:penta) (6 mL;
10 mg/mL).
Stable for > 5 years at -20°C.
Bottle 2: α-D-Glucuronidase solution (2 mL; ~ 1.0 U/mL, see
label on bottle).
Stable for > 2 years at -20°C.
Bottle 3: Stop Buffer (28 mL, pH 10) plus sodium azide
(0.02 % w/v) as a preservative.
Stable for > 2 years at 4°C.
Bottle 4: NAD+ freeze dried powder.
Stable for > 5 years at -20°C.
Bottle 5: Uronate dehydrogenase suspension (1.1 mL).
Stable for > 2 years at 4°C.