Cloning and expression of ATP N-glycosidase from the freshwater sponge Ephydatia muelleri

Biochimie. 2019 Mar:158:126-129. doi: 10.1016/j.biochi.2018.12.018. Epub 2018 Dec 29.

Abstract

Previously we had discovered unusual enzymatic activity in the marine sponge Axinella polypoides, ATP N-glycosidase (Reintamm et al., 2003). We show here that the Ephydatia muelleri mRNA encoding protein with PNP_UDP_1 (phosphorylase superfamily) signature is the secreted ATP N-glycosidase. The functionality of the protein was established by recombinant expression in Pichia pastoris. In addition to the enzymatic domain, the full-length protein contains the N-terminal cysteine-rich domain belonging to the subfamily SCP_HrTT-1 (cd05559) of the SCP (sperm coating protein) superfamily (cl00133).

Keywords: ATP N-Glycosidase; Ephydatia muelleri; PNP_UDP_1 domain; Pichia pastoris expression system; Porifera.

MeSH terms

  • Animals
  • Cloning, Molecular*
  • Gene Expression*
  • Glycoside Hydrolases* / biosynthesis
  • Glycoside Hydrolases* / chemistry
  • Glycoside Hydrolases* / classification
  • Glycoside Hydrolases* / genetics
  • Pichia / genetics
  • Pichia / metabolism
  • Porifera* / enzymology
  • Porifera* / genetics
  • Protein Domains
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / classification
  • Recombinant Proteins / genetics

Substances

  • Recombinant Proteins
  • Glycoside Hydrolases