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

ABSTRACT: AmyP is a raw-starch-degrading ?-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 ?-cyclodextrin. X-ray diffraction data were collected to a resolution of 2.1?Å. The crystal belonged to space group P2?2?2, with unit-cell parameters a=129.824, b=215.534, c=79.699?Å, ?=?=?=90°, and was estimated to contain two molecules in one asymmetric unit.

SUBMITTER: Yu J

PROVIDER: S-EPMC3606570 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature

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

Yu Jigang J Wang Chengliang C Hu Yanjin Y Dong Yuanqiu Y Wang Ying Y Tu Xiaoming X Peng Hui H Zhang Xuecheng X

Acta crystallographica. Section F, Structural biology and crystallization communications 20130222 Pt 3

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. ...[more]

PMID: 23519800

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