9BWP

Crystal structure of polyketoacyl-CoA thiolase from Burkholderia sp. in complex with acetoacetyl-coA

これはPDB形式変換不可エントリーです。

9BWP の概要

• エントリーDOI: 10.2210/pdb9bwp/pdb
• 分子名称: Acetyl-CoA acetyltransferase, COENZYME A, acetoacetyl group, ... (4 entities in total)
• 機能のキーワード: thiolases biosynthetic thiolases type iii polyketide synthase, lyase
• 由来する生物種: Burkholderia sp. RF2-non_BP3
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 84451.48

構造登録者

Pereira, J.H.,Wang, Z.,Keasling, J.D.,Adams, P.D. (登録日: 2024-05-21, 公開日: 2025-08-27, 最終更新日: 2025-12-24)

主引用文献

Wang, Z.,Cheong, S.,Pereira, J.H.,Hu, W.,Guo, Y.,DeGiovanni, A.,Lan, G.,Kim, J.,Haushalter, R.W.,Lee, T.S.,Adams, P.D.,Keasling, J.D.
A highly active Burkholderia polyketoacyl-CoA thiolase for production of triacetic acid lactone.
Nat Commun, 16:10990-10990, 2025

PubMed Abstract: Triacetic acid lactone (TAL) is a versatile platform chemical traditionally biosynthesized via decarboxylative Claisen condensation by 2-pyrone synthase. However, this route is limited by poor efficiency and dependence on malonyl-CoA. Here, we show that non-decarboxylative Claisen condensation by polyketoacyl-CoA thiolases offers a more efficient alternative. Through mining homologs of a previously reported enzyme from Cupriavidus necator, we identify five thiolases with TAL production activity. One candidate, BktBbr from Burkholderia sp. RF2-non_BP3, exhibits approximately 30-fold higher activity in vitro and supports 30-fold higher TAL titers in Escherichia coli compared to the original enzyme. Fed-batch fermentation achieves titers up to 2.8 g L⁻¹. Structural analysis of BktBbr co-crystallized with CoA esters guides rational engineering to further enhance performance. Our discovery of a highly active thiolase establishes an alternative enzymatic route to produce TAL efficiently, providing a scalable foundation for sustainable biomanufacturing.

PubMed: 41365852
DOI: 10.1038/s41467-025-65946-y

実験手法

X-RAY DIFFRACTION (2.45 Å)