TM1385 from Thermotoga maritima functions as a phosphoglucose isomerase via cis-enediol-based mechanism with active site redundancy

Swope, Nicole; Lake, Katherine E.; Barrow, Golda H.; Yu, Daniel; Fox, Daniel A.; Columbus, Linda

doi:10.1016/j.bbapap.2021.140602

Cited by 1 publication

(1 citation statement)

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“…However, the hydride shift mechanism in cupin phosphoglucose isomerases is still a matter of debate with experiments using NMR and EPR adding evidence to support the cis-enediol intermediate mechanism (Berrisford et al, 2006) while quantum mechanics/molecular mechanics studies supporting the hydride shift (Wu et al, 2008). For the conventional metal independent phosphoglucose isomerases, it has been well established that these enzymes operate by the cisenediol intermediate mechanism (Read et al, 2001;Swope et al, 2021).…”

Section: The Active Sitementioning

confidence: 99%

Biochemical and Structural Characterisation of a Novel D-Lyxose Isomerase From the Hyperthermophilic Archaeon Thermofilum sp.

Rose

Kuprat

Isupov

et al. 2021

Front. Bioeng. Biotechnol.

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A novel D-lyxose isomerase has been identified within the genome of a hyperthermophilic archaeon belonging to the Thermofilum species. The enzyme has been cloned and over-expressed in Escherichia coli and biochemically characterised. This enzyme differs from other enzymes of this class in that it is highly specific for the substrate D-lyxose, showing less than 2% activity towards mannose and other substrates reported for lyxose isomerases. This is the most thermoactive and thermostable lyxose isomerase reported to date, showing activity above 95°C and retaining 60% of its activity after 60 min incubation at 80°C. This lyxose isomerase is stable in the presence of 50% (v/v) of solvents ethanol, methanol, acetonitrile and DMSO. The crystal structure of the enzyme has been resolved to 1.4–1.7 A. resolution in the ligand-free form and in complexes with both of the slowly reacting sugar substrates mannose and fructose. This thermophilic lyxose isomerase is stabilised by a disulfide bond between the two monomers of the dimeric enzyme and increased hydrophobicity at the dimer interface. These overall properties of high substrate specificity, thermostability and solvent tolerance make this lyxose isomerase enzyme a good candidate for potential industrial applications.

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