3WPZ

Microbacterium saccharophilum K-1 beta-fructofuranosidase mutant T47S/S200T/F447P/F470Y/P500S

3WPZ の概要

• エントリーDOI: 10.2210/pdb3wpz/pdb
• 関連するPDBエントリー: 3VSR 3VSS 3WPU 3WPV 3WPY
• 分子名称: Beta-fructofuranosidase (2 entities in total)
• 機能のキーワード: glycoside hydrolase family 68, beta-propeller, hydrolase
• 由来する生物種: Arthrobacter
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 178131.57

構造登録者

Yokoi, G.,Mori, M.,Sato, S.,Miyazaki, T.,Nishikawa, A.,Tonozuka, T. (登録日: 2014-01-17, 公開日: 2014-03-12, 最終更新日: 2024-11-13)

主引用文献

Ohta, Y.,Hatada, Y.,Hidaka, Y.,Shimane, Y.,Usui, K.,Ito, T.,Fujita, K.,Yokoi, G.,Mori, M.,Sato, S.,Miyazaki, T.,Nishikawa, A.,Tonozuka, T.
Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 beta-fructofuranosidase
Appl.Microbiol.Biotechnol., 98:6667-6677, 2014

PubMed Abstract: A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose (4(G)-β-D-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes.

PubMed: 24633372
DOI: 10.1007/s00253-014-5645-3

実験手法

X-RAY DIFFRACTION (2.27 Å)