Purification, crystallization and preliminary crystallographic analysis of the marine α-amylase AmyP

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Mar 1;69(Pt 3):263-6. doi: 10.1107/S1744309113001693. Epub 2013 Feb 22.

Abstract

AmyP is a raw-starch-degrading α-amylase newly identified from a marine metagenome library. It shares low sequence similarity with characterized glycoside hydrolases and was classified into a new subfamily of GH13. In particular, it showed preferential degradation to raw rice starch. Full-length AmyP was cloned and overexpressed in Escherichia coli, then purified and crystallized in the presence of its substrate analogue β-cyclodextrin. X-ray diffraction data were collected to a resolution of 2.1 Å. The crystal belonged to space group P2₁2₁2, with unit-cell parameters a=129.824, b=215.534, c=79.699 Å, α=β=γ=90°, and was estimated to contain two molecules in one asymmetric unit.

Keywords: AmyP; α-amylase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aquatic Organisms / chemistry*
  • Aquatic Organisms / enzymology
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / chemistry
  • Escherichia coli / genetics
  • Metagenome*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Starch / chemistry
  • Starch / metabolism
  • alpha-Amylases / chemistry*
  • alpha-Amylases / genetics
  • beta-Cyclodextrins / chemistry*

Substances

  • Recombinant Proteins
  • beta-Cyclodextrins
  • Starch
  • alpha-Amylases
  • betadex