8I07

Crystal structure of Escherichia coli glyoxylate carboligase double mutant in complex with glycolaldehyde

Summary for 8I07

• Entry DOI: 10.2210/pdb8i07/pdb
• Related: 8I01 8I05
• Descriptor: Glyoxylate carboligase, MAGNESIUM ION, FLAVIN-ADENINE DINUCLEOTIDE, ... (7 entities in total)
• Functional Keywords: ligase, gcl
• Biological source: Escherichia coli K-12
• Total number of polymer chains: 6
• Total formula weight: 399957.94

Authors

Kim, J.H.,Kim, J.S. (deposition date: 2023-01-10, release date: 2023-11-22, Last modification date: 2024-05-08)

Primary citation

Kim, J.H.,Cheon, H.,Jo, H.J.,Kim, J.W.,Kim, G.Y.,Seo, H.R.,Seo, P.W.,Kim, J.S.,Park, J.B.
Engineering of two thiamine diphosphate-dependent enzymes for the regioselective condensation of C1-formaldehyde into C4-erythrulose.
Int.J.Biol.Macromol., 253:127674-127674, 2023

PubMed Abstract: A number of carboligases, which catalyze condensation of C1- and/or C2-aldehydes into multi-carbon products, have been reported. However, their catalytic activities and/or regioselectivities remained rather low. Thereby, this study has focused on engineering of C1 and C2 carboligases for the regioselective condensation of C1-formaldehyde into C4-erythrulose via C2-glycolaldehyde. The crystal structure of the glyoxylate carboligase from Escherichia coli (EcGCL) was elucidated in complex with glycolaldehyde. A structure-guided rationale generated several mutants, one of whose catalytic activity reached 15.6 M·s, almost 10 times greater than the wild-type enzyme. Another variant (i.e., EcGCL) has shown significantly increased stability to the glycolaldehyde toxicity, enabling production of glycolaldehyde to 31 mM from 75 mM formaldehyde (conversion: 83 %). Besides, the E1 subunit of α-ketoglutarate dehydrogenase complex from Vibrio vulnificus (VvSucA) was engineered as a regiospecific C2 carboligase for condensation of glycolaldehyde into erythrulose. The combination of EcGCL and VvSucA led to the cascade production of erythrulose to 8 mM from 90 mM formaldehyde via glycolaldehyde without byproduct formation. This study will contribute to valorization of C1 gases into industrially relevant multi-carbon products in an environment-friendly way.

PubMed: 37890751
DOI: 10.1016/j.ijbiomac.2023.127674

Experimental method

X-RAY DIFFRACTION (1.99 Å)