7B2E

quadruple mutant of oxalyl-CoA decarboxylase from Methylorubrum extorquens with bound TPP and ADP

7B2E の概要

• エントリーDOI: 10.2210/pdb7b2e/pdb
• 関連するPDBエントリー: 7AYG
• 分子名称: Putative oxalyl-CoA decarboxylase (Oxc, yfdU), THIAMINE DIPHOSPHATE, ADENOSINE-5'-DIPHOSPHATE, ... (5 entities in total)
• 機能のキーワード: lyase, decarboxylase, carboxylase, ligase, formyl-coa, oxalyl-coa, mandelyl-coa
• 由来する生物種: Methylorubrum extorquens (strain ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1)
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 500477.46

構造登録者

Pfister, P.,Burgener, S.,Nattermann, M.,Zarzycki, J.,Erb, T.J. (登録日: 2020-11-26, 公開日: 2021-04-28, 最終更新日: 2024-01-31)

主引用文献

Nattermann, M.,Burgener, S.,Pfister, P.,Chou, A.,Schulz, L.,Lee, S.H.,Paczia, N.,Zarzycki, J.,Gonzalez, R.,Erb, T.J.
Engineering a Highly Efficient Carboligase for Synthetic One-Carbon Metabolism.
Acs Catalysis, 11:5396-5404, 2021

PubMed Abstract: One of the biggest challenges to realize a circular carbon economy is the synthesis of complex carbon compounds from one-carbon (C1) building blocks. Since the natural solution space of C1-C1 condensations is limited to highly complex enzymes, the development of more simple and robust biocatalysts may facilitate the engineering of C1 assimilation routes. Thiamine diphosphate-dependent enzymes harbor great potential for this task, due to their ability to create C-C bonds. Here, we employed structure-guided iterative saturation mutagenesis to convert oxalyl-CoA decarboxylase (OXC) from into a glycolyl-CoA synthase (GCS) that allows for the direct condensation of the two C1 units formyl-CoA and formaldehyde. A quadruple variant MeOXC4 showed a 100 000-fold switch between OXC and GCS activities, a 200-fold increase in the GCS activity compared to the wild type, and formaldehyde affinity that is comparable to natural formaldehyde-converting enzymes. Notably, MeOCX4 outcompetes all other natural and engineered enzymes for C1-C1 condensations by more than 40-fold in catalytic efficiency and is highly soluble in . In addition to the increased GCS activity, MeOXC4 showed up to 300-fold higher activity than the wild type toward a broad range of carbonyl acceptor substrates. When applied in vivo, MeOXC4 enables the production of glycolate from formaldehyde, overcoming the current bottleneck of C1-C1 condensation in whole-cell bioconversions and paving the way toward synthetic C1 assimilation routes in vivo.

PubMed: 34484855
DOI: 10.1021/acscatal.1c01237

実験手法

X-RAY DIFFRACTION (2.8 Å)