A bacterial endo-β-1,4-glucuronan lyase, CUL-I from Brevundimonas sp. SH203, belonging to a novel polysaccharide lyase family

Protein Expr Purif. 2020 Feb:166:105502. doi: 10.1016/j.pep.2019.105502. Epub 2019 Sep 20.

Abstract

Cellouronate is a (1,4)-β-D-glucuronan prepared by TEMPO-mediated oxidation from regenerated cellulose. We have previously isolated a cellouronate-degrading bacterial strain, Brevundimonas sp. SH203, that produces a cellouronate lyase (β-1,4-glucuronan lyase, CUL-I). In this study, the gene encoding CUL-I was cloned, and the recombinant enzyme was heterologously expressed in Escherichia coli. The predicted CUL-I protein is composed of 426 amino acid residues and includes a putative 21-amino acid signal peptide. The recombinant CUL-I specifically depolymerized β-1,4-glycoside linkages of cellouronate, and its mode of action was endo-type, like the native CUL-I. Sequence analysis showed CUL-I has no similarity to previously known polysaccharide lyases (PLs), indicating that CUL-I should be classified into a novel PL family.

Keywords: Brevundimonas sp; Cellouronate; Polysaccharide lyase (PL); TEMPO-Mediated oxidation; β-1,4-Glucuronan lyase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Caulobacteraceae / enzymology
  • Caulobacteraceae / genetics*
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Gene Expression
  • Glycosides / chemistry
  • Glycosides / metabolism
  • Oxidation-Reduction
  • Polysaccharide-Lyases / chemistry
  • Polysaccharide-Lyases / classification
  • Polysaccharide-Lyases / genetics*
  • Protein Sorting Signals / genetics
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / classification
  • Recombinant Proteins / genetics*

Substances

  • Glycosides
  • Protein Sorting Signals
  • Recombinant Proteins
  • Polysaccharide-Lyases
  • glucuronan lyase