9QYU

Crystal structure of leaf branch compost cutinase quintuple variant ICCG L50Y

9QYU の概要

• エントリーDOI: 10.2210/pdb9qyu/pdb
• 分子名称: Leaf-branch compost cutinase, 1,2-ETHANEDIOL, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: hydrolase, cutinase, pet depolymerase
• 由来する生物種: uncultured bacterium
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 27993.72

構造登録者

Bischoff, D.,Walla, B.,Dietrich, A.-M.,Janowski, R.,Niessing, D.,Weuster-Botz, D. (登録日: 2025-04-21, 公開日: 2025-07-02, 最終更新日: 2025-07-09)

主引用文献

Walla, B.,Dietrich, A.M.,Brames, E.,Bischoff, D.,Fritzsche, S.,Castiglione, K.,Janowski, R.,Niessing, D.,Weuster-Botz, D.
Application of a Rational Crystal Contact Engineering Strategy on a Poly(ethylene terephthalate)-Degrading Cutinase.
Bioengineering (Basel), 12:-, 2025

PubMed Abstract: Industrial biotechnology offers a potential ecological solution for PET recycling under relatively mild reaction conditions via enzymatic degradation, particularly using the leaf branch compost cutinase (LCC) quadruple mutant ICCG. To improve the efficient downstream processing of this biocatalyst after heterologous gene expression with a suitable production host, protein crystallization can serve as an effective purification/capture step. Enhancing protein crystallization was achieved in recent studies by introducing electrostatic (and aromatic) interactions in two homologous alcohol dehydrogenases (/ADH) and an ene reductase (ER1-L1,5) produced with . In this study, ICCG, which is difficult to crystallize, was utilized for the application of crystal contact engineering strategies, resulting in ICCG mutant L50Y (ICCGY). Previously focused on the Lys-Glu interaction for the introduction of electrostatic interactions at crystal contacts, the applicability of the engineering strategy was extended here to an Arg-Glu interaction to increase crystallizability, as shown for ICCGY T110E. Furthermore, the rationale of the engineering approach is demonstrated by introducing Lys and Glu at non-crystal contacts or sites without potential interaction partners as negative controls. These resulting mutants crystallized comparably but not superior to the wild-type protein. As demonstrated by this study, crystal contact engineering emerges as a promising approach for rationally enhancing protein crystallization. This advancement could significantly streamline biotechnological downstream processing, offering a more efficient pathway for research and industry.

PubMed: 40564377
DOI: 10.3390/bioengineering12060561

実験手法

X-RAY DIFFRACTION (1.51 Å)