8A85

Structure of the Reconstructed Ancestor of Phenolic Acid Decarboxylase AncPAD134

8A85 の概要

• エントリーDOI: 10.2210/pdb8a85/pdb
• 分子名称: Phenolic acid decarboxylase N134 (2 entities in total)
• 機能のキーワード: lyase, phenolic acid decarboxylase, decarboxylase, phenolic-acid, ancestral sequence, decarboxylation, ancestor, n134, bacillus subtilis
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 128837.66

構造登録者

Mokos, D.,Schruefer, A.,Gruber, K.,Daniel, B. (登録日: 2022-06-22, 公開日: 2023-07-05, 最終更新日: 2025-01-15)

主引用文献

Myrtollari, K.,Calderini, E.,Kracher, D.,Schongassner, T.,Galusic, S.,Slavica, A.,Taden, A.,Mokos, D.,Schrufer, A.,Wirnsberger, G.,Gruber, K.,Daniel, B.,Kourist, R.
Stability Increase of Phenolic Acid Decarboxylase by a Combination of Protein and Solvent Engineering Unlocks Applications at Elevated Temperatures.
Acs Sustain Chem Eng, 12:3575-3584, 2024

PubMed Abstract: Enzymatic decarboxylation of biobased hydroxycinnamic acids gives access to phenolic styrenes for adhesive production. Phenolic acid decarboxylases are proficient enzymes that have been applied in aqueous systems, organic solvents, biphasic systems, and deep eutectic solvents, which makes stability a key feature. Stabilization of the enzyme would increase the total turnover number and thus reduce the energy consumption and waste accumulation associated with biocatalyst production. In this study, we used ancestral sequence reconstruction to generate thermostable decarboxylases. Investigation of a set of 16 ancestors resulted in the identification of a variant with an unfolding temperature of 78.1 °C and a half-life time of 45 h at 60 °C. Crystal structures were determined for three selected ancestors. Structural attributes were calculated to fit different regression models for predicting the thermal stability of variants that have not yet been experimentally explored. The models rely on hydrophobic clusters, salt bridges, hydrogen bonds, and surface properties and can identify more stable proteins out of a pool of candidates. Further stabilization was achieved by the application of mixtures of natural deep eutectic solvents and buffers. Our approach is a straightforward option for enhancing the industrial application of the decarboxylation process.

PubMed: 38456190
DOI: 10.1021/acssuschemeng.3c06513

実験手法

X-RAY DIFFRACTION (2.67 Å)