7FIP

The native structure of beta-1,2-mannobiose phosphorylase from Thermoanaerobacter sp.

7FIP の概要

• エントリーDOI: 10.2210/pdb7fip/pdb
• 分子名称: Beta-1,2-mannobiose phosphorylase, ZINC ION (3 entities in total)
• 機能のキーワード: mannobiose, phosphorylase, thermoanaerobacter, transferase, crystallization, complex structure
• 由来する生物種: Thermoanaerobacter sp. (strain X514)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 144474.15

構造登録者

Dai, L.,Chang, Z.,Yang, J.,Liu, W.,Yang, Y.,Chen, C.-C.,Zhang, L.,Huang, J.,Sun, Y.,Guo, R.-T. (登録日: 2021-08-01, 公開日: 2022-01-05, 最終更新日: 2023-11-29)

主引用文献

Dai, L.,Chang, Z.,Yang, J.,Liu, W.,Yang, Y.,Chen, C.C.,Zhang, L.,Huang, J.W.,Sun, Y.,Guo, R.T.
Structural investigation of a thermostable 1,2-beta-mannobiose phosphorylase from Thermoanaerobacter sp. X-514.
Biochem.Biophys.Res.Commun., 579:54-61, 2021

PubMed Abstract: 1,2-β-Mannobiose phosphorylases (1,2-β-MBPs) from glycoside hydrolase 130 (GH130) family are important bio-catalysts in glycochemistry applications owing to their ability in synthesizing oligomannans. Here, we report the crystal structure of a thermostable 1,2-β-MBP from Thermoanaerobacter sp. X-514 termed Teth514_1789 to reveal the molecular basis of its higher thermostability and mechanism of action. We also solved the enzyme complexes of mannose, mannose-1-phosphate (M1P) and 1,4-β-mannobiose to manifest the enzyme-substrate interaction networks of three main subsites. Notably, a Zn ion that should be derived from crystallization buffer was found in the active site and coordinates the phosphate moiety of M1P. Nonetheless, this Zn-coordination should reflect an inhibitory status as supplementing Zn severely impairs the enzyme activity. These results indicate that the effects of metal ions should be taken into consideration when applying Teth514_1789 and other related enzymes. Based on the structure, a reliable model of Teth514_1788 that shares 61.7% sequence identity to Teth514_1789 but displays a different substrate preference was built. Analyzing the structural features of these two closely related enzymes, we hypothesized that the length of a loop fragment that covers the entrance of the catalytic center might regulate the substrate selectivity. In conclusion, these information provide in-depth understanding of GH130 1,2-β-MBPs and should serve as an important guidance for enzyme engineering for further applications.

PubMed: 34587555
DOI: 10.1016/j.bbrc.2021.09.046

実験手法

X-RAY DIFFRACTION (2.39 Å)