Crystallization and preliminary X‐ray crystallographic analysis of the amylomaltase from Corynebacterium glutamicum. Issue 9 (30th August 2013)

• Record Type: Journal Article
• Authors: Srisimarat, Wiraya; Murakami, Shuichiro; Pongsawasdi, Piamsook; Krusong, Kuakarun
• Is Part Of: Acta crystallographica. Volume 69:Issue 9(2013:Sep.)
• Page Start: 1004
• Page End: 1006
• Publication Date: 2013-08-30
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S1744309113020319
• Languages: English
• ISSNs: 1744-3091

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Amylomaltase (AM; EC 2.4.1.25) belongs to the 4‐α‐glucanotransferase group of the α‐amylase family. The enzyme can produce cycloamylose or large‐ring cyclodextrin through intramolecular transglycosylation or cyclization reactions of α‐1, 4‐glucan. Amylomaltase from the mesophilic bacterium <italic>Corynebacterium glutamicum</italic> (<italic>Cg</italic>AM) contains extra residues at the N‐terminus for which the three‐dimensional structure is not yet known. In this study, <italic>Cg</italic>AM was overexpressed and purified to homogeneity using DEAE FF and Phenyl FF columns. The purified <italic>Cg</italic>AM was crystallized by the vapour‐diffusion method. Preliminary X‐ray data showed that the <italic>Cg</italic>AM crystal diffracted to 1.7 Å resolution and belonged to space group <italic>P</italic>2₁2₁2₁, with unit‐cell parameters <italic>a</italic> = 73.28, <italic>b</italic> = 82.61, <italic>c</italic> = 118.64 Å. To obtain the initial phases, crystals of selenomethionyl‐substituted amylomaltase were produced, and multiple‐wavelength anomalous dispersion phasing and structure refinement are now in progress.</p> </abstract>