Isolation of α-Glucuronidase Enzyme from a Rumen Metagenomic Library
- 302 Downloads
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.
Keywordsα-Glucuronidase Rumen Metagenome Library screening Hemicellulose
Open reading frame
α-Glucuronidase from rumen microorganism
Mention of trade names or commercial products in this report is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer.
- 1.Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) J Mol Biol 215:403–410Google Scholar
- 2.Biely P (2002) In: Whitaker JR, Voragen AG, Wong DWS (eds) Handbook of food enzymology. Marcel Dekker, New York, NY, pp 879–915Google Scholar
- 9.Henrissat B, Bairoch A (1996) Biochem J 316:695–696Google Scholar
- 11.Iiyama K, Lam T, Stone BA (1994) Plant Physiol 104:315–320Google Scholar
- 28.Shulami S, Gat O, Sonenshein AL, Shoham Y (1999) J Bacteriol 181:3695–3704Google Scholar
- 31.Spiridon I, Popa VI (2005) In: Dumitriu S (ed) Polysaccharides: structural diversity and functional versatility, 2nd edn. Marcel Dekker, New York, NY, pp 475–489Google Scholar
- 35.Timell TE (1964) Adv Carbohydr Chem 19:247–302Google Scholar
- 36.Timell TE (1965) Adv Carbohydr Chem 20:409–483Google Scholar