Crystallization and preliminary X-ray analysis of a novel halotolerant feruloyl esterase identified from a soil metagenomic library

Abstract

Feruloyl esterase cleaves the ester linkage formed between ferulic acid and polysaccharides in plant cell walls and thus has wide potential industrial applications. A novel feruloyl esterase (EstF27) identified from a soil metagenomic library was crystallized and a complete data set was collected from a single cooled crystal using an in-house X-ray source. The crystal diffracted to 2.9 Å resolution and belonged to space group P2 1 2 1 2 1 , with unitcell parameters a = 94.35, b = 106.19, c = 188.51 Å , = = = 90.00 . A Matthews coefficient of 2.55 Å 3 Da À1 , with a corresponding solvent content of 51.84%, suggested the presence of ten protein subunits in the asymmetric unit.

Enhancing the Thermostability of Feruloyl Esterase EstF27 by Directed Evolution and the Underlying Structural Basis

“…The unit cell was relatively large, and 10 protein subunits were likely to be present in the asymmetric unit. 32 Molecular replacement using various available structures was not successful. Consequently, we solved the structure of M6 using experimental phasing with CsCl as the anomalous scatterer.…”

“…The crystal of the WT was obtained from a solution containing PEG 8000. The unit cell was relatively large, and 10 protein subunits were likely to be present in the asymmetric unit . Molecular replacement using various available structures was not successful.…”