3WFZ

Crystal structure of Galacto-N-Biose/Lacto-N-Biose I Phosphorylase C236Y Mutant

3WFZ の概要

• エントリーDOI: 10.2210/pdb3wfz/pdb
• 関連するPDBエントリー: 2ZUU
• 分子名称: Lacto-N-biose phosphorylase, 2-acetamido-2-deoxy-alpha-D-glucopyranose (3 entities in total)
• 機能のキーワード: beta-alpha-barrel, tim barrel, phosphorylase, transferase
• 由来する生物種: Bifidobacterium longum
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 343061.21

構造登録者

Koyama, Y.,Hidaka, M.,Kawakami, M.,Nishimoto, M.,Kitaoka, M. (登録日: 2013-07-25, 公開日: 2013-10-09, 最終更新日: 2023-11-08)

主引用文献

Koyama, Y.,Hidaka, M.,Nishimoto, M.,Kitaoka, M.
Directed evolution to enhance thermostability of galacto-N-biose/lacto-N-biose I phosphorylase.
Protein Eng.Des.Sel., 26:755-761, 2013

PubMed Abstract: Galacto-N-biose/lacto-N-biose I phosphorylase (GLNBP) is the key enzyme in the enzymatic production of lacto-N-biose I. For the purpose of industrial use, we improved the thermostability of GLNBP by evolutionary engineering in which five substitutions in the amino acid sequence were selected from a random mutagenesis GLNBP library constructed using error-prone polymerase chain reaction. Among them, C236Y and D576V mutants showed considerably improved thermostability. Structural analysis of C236Y revealed that the hydroxyl group of Tyr236 forms a hydrogen bond with the carboxyl group of E319. The C236Y and D576V mutations together contributed to the thermostability. The C236Y/D576V mutant exhibited 20°C higher thermostability than the wild type.

PubMed: 24065834
DOI: 10.1093/protein/gzt049

実験手法

X-RAY DIFFRACTION (2.6 Å)