4AEF

THE CRYSTAL STRUCTURE OF THERMOSTABLE AMYLASE FROM THE PYROCOCCUS

4AEF の概要

• エントリーDOI: 10.2210/pdb4aef/pdb
• 分子名称: NEOPULLULANASE (ALPHA-AMYLASE II) (2 entities in total)
• 機能のキーワード: hydrolase, thermostability, high temperature
• 由来する生物種: PYROCOCCUS FURIOSUS
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 152391.64

構造登録者

Song, H.-N.,Jung, T.-Y.,Yoon, S.-M.,Yang, S.-J.,Park, K.-H.,Woo, E.-J. (登録日: 2012-01-10, 公開日: 2012-10-31, 最終更新日: 2023-12-20)

主引用文献

Park, J.-T.,Song, H.-N.,Jung, T.-Y.,Lee, M.H.,Park, S.G.,Woo, E.-J.,Park, K.-H.
A Novel Domain Arrangement in a Monomeric Cyclodextrin-Hydrolyzing Enzyme from the Hyperthermophile Pyrococcus Furiosus.
Biochim.Biophys.Acta, 1834:380-, 2013

PubMed Abstract: PFTA (Pyrococcus furiosus thermostable amylase) is a hyperthermophilic amylase isolated from the archaeon Pyrococcus furiosus. This enzyme possesses characteristics of both α-amylase- and cyclodextrin (CD)-hydrolyzing enzymes, allowing it to degrade pullulan, CD and acarbose-activities that are absent in most α-amylases-without the transferring activity that is common in CD-hydrolyzing enzymes. The crystal structure of PFTA revealed a unique monomeric subunit with an extended N-terminal region and an N'-domain folded into its own active site-a significantly altered domain configuration relative to that of the conventional dimeric CD-hydrolyzing amylases in glycoside hydrolase family 13. The active site is formed by the interface of the N'-domain and the catalytic domain and exhibits a broad and wide-open geometry without the concave pocket that is commonly found in the active sites of maltogenic amylases. The mutation of a residue (Gly415 to Glu) located at the domain interface between the N'- and catalytic domains yielded an enzyme that produced a significantly higher purity maltoheptaose (G7) from β-CD, supporting the involvement of this interface in substrate recognition and indicating that this mutant enzyme is a suitable candidate for the production of pure G7. The unique configuration of the active site distinguishes this archaic monomeric enzyme from classical bacterial CD-hydrolyzing amylases and provides a molecular basis for its enzymatic characteristics and for its potential use in industrial applications.

PubMed: 22902546
DOI: 10.1016/J.BBAPAP.2012.08.001

実験手法

X-RAY DIFFRACTION (2.34 Å)