8B30
Structure of the Reconstructed Ancestor of Phenolic Acid Decarboxylase AncPAD31
Summary for 8B30
| Entry DOI | 10.2210/pdb8b30/pdb |
| Related | 8A85 |
| Descriptor | Phenolic acid decarboxylase N31 (2 entities in total) |
| Functional Keywords | lyase, phenolic acid decarboxylase, decarboxylase, phenolic-acid, ancestral sequence, decarboxylation, ancestor, n31, bacillus subtilis |
| Biological source | synthetic construct |
| Total number of polymer chains | 2 |
| Total formula weight | 43799.02 |
| Authors | Mokos, D.,Schruefer, A.,Gruber, K.,Daniel, B. (deposition date: 2022-09-15, release date: 2023-09-27, Last modification date: 2024-03-20) |
| Primary citation | 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 Cited by 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: 38456190DOI: 10.1021/acssuschemeng.3c06513 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.7 Å) |
Structure validation
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