Isolation of α-glucuronidase enzyme from a rumen metagenomic library.
α-Glucuronidase enzymes play an essential role in the full enzymatic hydrolysis of hemicellulose. Up to this point, all genes encoding α-glucuronidase enzymes have been cloned from individual, pure culture strains. Using a high-throughput screening strategy, we have isolated the first α-glucuronidase gene (rum630-AG) from a mixed population of microorganisms. The gene was subcloned into a prokaryotic vector, and the enzyme was overexpressed and biochemically characterized. The RUM630-AG enzyme had optimum activity at pH 6.5 and 40 °C. When birchwood xylan was used as substrate, the RUM630-AG functioned synergistically with an endoxylanase enzyme to hydrolyze the substrate.
USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710, USA. Charles.Lee@ars.usda.gov
Charles C
CC
Lee
Rena E
RE
Kibblewhite
Kurt
K
Wagschal
Ruiping
R
Li
William J
WJ
Orts
D000595:Amino Acid Sequence; D000818:Animals; D029662:Betula; D002417:Cattle; D003001:Cloning, Molecular; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D015698:Genomic Library; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D056186:Metagenomics / Q000379:methods*; D008969:Molecular Sequence Data; D012417:Rumen / Q000201:enzymology* / Q000382:microbiology; D016415:Sequence Alignment; D014990:Xylans / Q000378:metabolism
D014990:Xylans; D006026:Glycoside Hydrolases; C096734:alpha-glucuronidase; D043364:Endo-1,4-beta Xylanases