4MOO

Pyranose 2-oxidase H450G mutant with 2-fluorinated galactose

4MOO の概要

• エントリーDOI: 10.2210/pdb4moo/pdb
• 関連するPDBエントリー: 1TT0 2IGO 3PL8 4MOE 4MOF 4MOG 4MOH 4MOI 4MOJ 4MOK 4MOL 4MOM 4MOP 4MOQ 4MOR 4MOS
• 分子名称: Pyranose 2-oxidase, DIHYDROFLAVINE-ADENINE DINUCLEOTIDE, 2-deoxy-2-fluoro-alpha-D-galactopyranose, ... (4 entities in total)
• 機能のキーワード: gmc oxidoreductase, phbh fold, homotetramer, fad-binding, substrate complex, flavinylation, intracellular, oxidoreductase
• 由来する生物種: Trametes ochracea (White-rot fungus)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 71442.67

構造登録者

Tan, T.C.,Spadiut, O.,Gandini, R.,Haltrich, D.,Divne, C. (登録日: 2013-09-12, 公開日: 2014-02-05, 最終更新日: 2024-10-30)

主引用文献

Tan, T.C.,Spadiut, O.,Gandini, R.,Haltrich, D.,Divne, C.
Structural Basis for Binding of Fluorinated Glucose and Galactose to Trametes multicolor Pyranose 2-Oxidase Variants with Improved Galactose Conversion.
Plos One, 9:e86736-e86736, 2014

PubMed Abstract: Each year, about six million tons of lactose are generated from liquid whey as industrial byproduct, and optimally this large carbohydrate waste should be used for the production of value-added products. Trametes multicolor pyranose 2-oxidase (TmP2O) catalyzes the oxidation of various monosaccharides to the corresponding 2-keto sugars. Thus, a potential use of TmP2O is to convert the products from lactose hydrolysis, D-glucose and D-galactose, to more valuable products such as tagatose. Oxidation of glucose is however strongly favored over galactose, and oxidation of both substrates at more equal rates is desirable. Characterization of TmP2O variants (H450G, V546C, H450G/V546C) with improved D-galactose conversion has been given earlier, of which H450G displayed the best relative conversion between the substrates. To rationalize the changes in conversion rates, we have analyzed high-resolution crystal structures of the aforementioned mutants with bound 2- and 3-fluorinated glucose and galactose. Binding of glucose and galactose in the productive 2-oxidation binding mode is nearly identical in all mutants, suggesting that this binding mode is essentially unaffected by the mutations. For the competing glucose binding mode, enzyme variants carrying the H450G replacement stabilize glucose as the α-anomer in position for 3-oxidation. The backbone relaxation at position 450 allows the substrate-binding loop to fold tightly around the ligand. V546C however stabilize glucose as the β-anomer using an open loop conformation. Improved binding of galactose is enabled by subtle relaxation effects at key active-site backbone positions. The competing binding mode for galactose 2-oxidation by V546C stabilizes the β-anomer for oxidation at C1, whereas H450G variants stabilize the 3-oxidation binding mode of the galactose α-anomer. The present study provides a detailed description of binding modes that rationalize changes in the relative conversion rates of D-glucose and D-galactose and can be used to refine future enzyme designs for more efficient use of lactose-hydrolysis byproducts.

PubMed: 24466218
DOI: 10.1371/journal.pone.0086736

実験手法

X-RAY DIFFRACTION (1.65 Å)