Initially, aspartate 631 acts as a catalytic base, abstracting a proton from the alpha carbon of acetyl-CoA and creating an enolate that is stabilized by arginine 338.

[7] Then, the newly formed enolate acts as a nucleophile that attacks the aldehyde of glyoxylate, imparting a negative charge on the oxygen which is stabilized by arginine 338 and the coordinating magnesium cation.

The glyoxylate cycle, facilitated by malate synthase and isocitrate lyase, allows plants and bacteria to subsist on acetyl-CoA or other two carbon compounds.

CLYBL differs from other malate synthases in that it lacks a large portion of the C-terminal domain and shows lower specific activity and efficiency.

[14][15] Because the glyoxylate cycle occurs in bacteria and fungi, studying the mechanisms of malate synthase (as well as isocitrate lyase) is important for understanding human, animal, and plant pathogenesis.

Studying malate synthase can shed light on the metabolic pathways that allow pathogens to survive inside a host as well as elucidate possible treatments.

[16] Many studies have been conducted on malate synthase activity in pathogens, including Mycobacterium tuberculosis, Pseudomonas aeruginosa, Brucella melitensis, and Escherichia coli.

[19] Downregulation of malate synthase results in reduced stress tolerance, persistence, and growth of Mycobacterium tuberculosis inside macrophages.

[21] Pseudomonas aeruginosa causes severe infections in humans and is labeled as a critical threat by the World Health Organization because of its resistance to multiple therapies.

In 2017, McVey, et al. examined the 3D structure of P. aeruginosa malate synthase G. They found that it is a monomer composed of four domains and is highly conserved in other pathogens.

[22] Brucella melitensis is a pathogenic bacterium that causes fever and inflammation of the epididymis in sheep and cattle and can be transmitted to humans through the consumption of unpasteurized milk.

In 2016, Adi, et al. constructed a 3D crystallized structure of the protein to identify catalytic domains and investigate potential inhibitors.

[23] In Escherichia coli, the genes encoding the enzymes required for the glyoxylate cycle are expressed from the polycistronic ace operon.