The GH67 α-glucuronidase of Paenibacillus curdlanolyticus B-6 removes hexenuronic acid groups and facilitates biodegradation of the model xylooligosaccharide hexenuronosyl xylotriose

Abstract

4-O-Methylglucuronic acid (MeGlcA) side groups attached to the xylan backbone through α-1,2 linkages are converted to hexenuronic acid (HexA) during alkaline pulping. α-Glucuronidase (EC 3.2.1.139) hydrolyzes 1,2-linked MeGlcA from xylooligosaccharides. To determine whether α-glucuronidase can also hydrolyze HexA-decorated xylooligosaccharides, a gene encoding α-glucuronidase (AguA) was cloned from Paenibacillus curdlanolyticus B-6. The purified protein degraded hexenuronosyl xylotriose (ΔX3), a model substrate prepared from kraft pulp. AguA released xylotriose and HexA from ΔX3, but the Vmax and kcat values for ΔX3 were lower than those for MeGlcA, indicating that HexA side groups may affect the hydrolytic activity. To explore the potential for biological bleaching, ΔX3 degradation was performed using intracellular extract from P. curdlanolyticus B-6. The intracellular extract, with synergistic α-glucuronidase and β-xylosidase activities, degraded ΔX3 to xylose and HexA. These results indicate that α-glucuronidase can be used to remove HexA from ΔX3 derived from pulp, reducing the need for chemical treatments in the pulping process.

Keywords: GH67; Hexenuronic acid; Hexenuronosyl xylotriose; Paenibacillus curdlanolyticus; α-Glucuronidase.