A unique CE16 acetyl esterase from Podospora anserina active on polymeric xylan.

Abstract

The genome of the coprophilous fungus Podospora anserina displays an impressive array of genes encoding hemicellulolytic enzymes. In this study, we focused on a putative carbohydrate esterase (CE) from family 16 (CE16) that bears a carbohydrate-binding module from family CBM1. The protein was heterologously expressed in Pichia pastoris and purified to electrophoretic homogeneity. The P. anserina CE16 enzyme (PaCE16A) exhibited different catalytic properties than so far known CE16 esterases represented by the Trichoderma reesei CE16 acetyl esterase (TrCE16). A common property of both CE16 esterases is their exodeacetylase activity, i.e., deesterification at positions 3 and 4 of monomeric xylosides and the nonreducing end xylopyranosyl (Xylp) residue of oligomeric homologues. However, the PaCE16A showed lower positional specificity than TrCE16 and efficiently deacetylated also position 2. The major difference observed between PaCE16A and TrCE16 was found on polymeric substrate, acetylglucuronoxylan. While TrCE16 does not attack internal acetyl groups, PaCE16A deacetylated singly and doubly acetylated Xylp residues in the polymer to such an extent that it resulted in the polymer precipitation. Similarly as typical acetylxylan esterases belonging to CE1, CE4, CE5, and CE6 families, PaCE16A did not attack 3-O-acetyl group of xylopyranosyl residues carrying 4-O-methyl-D-glucuronic acid at position 2. PaCE16A thus represents a CE16 member displaying unique catalytic properties, which are intermediate between the TrCE16 exodeacetylase and acetylxylan esterases designed to deacetylate polymeric substrate. The catalytic versatility of PaCE16A makes the enzyme an important candidate for biotechnological applications.