Serine hydroxymethyltransferase (SHMT) and threonine aldolase are classified as fold type I pyridoxal-5’-phosphate-dependent enzymes and engaged in glycine biosynthesis from serine and threonine, respectively. The acidothermophilic archaeon Thermoplasma acidophilum possesses two distinct SHMT genes, while there is no gene encoding threonine aldolase in its genome. In the present study, the two SHMT genes (Ta0811 and Ta1509) were heterologously expressed in Escherichia coli and Thermococcus kodakarensis, respectively, and biochemical properties of their products were investigated. Ta1509 protein exhibited dual activities to catalyze tetrahydrofolate (THF)-dependent serine cleavage and THF-independent threonine cleavage, similar to other SHMTs reported to date. In contrast, the Ta0811 protein lacks amino acid residues involved in the THF-binding motif and catalyzes only the THF-independent cleavage of threonine. Kinetic analysis revealed that the threonine-cleavage activity of the Ta0811 protein was 3.5 times higher than the serine-cleavage activity of Ta1509 protein. In addition, mRNA expression of Ta0811 gene in T. acidophilum was approximately 20 times more abundant than that of Ta1509. These observations suggest that retroaldol cleavage of threonine, mediated by the Ta0811 protein, has a major role in glycine biosynthesis in T. acidophilum.
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This work was partly supported by the JSPS KAKENHI program (grant numbers 17K07720 and 20H05586). This work was also supported by a Grant-in-Aid for JSPS Fellows (Grant number 20J00010). We thank Dr. Tamotsu Kanai (Toyama Prefectural University) and Dr. Haruyuki Atomi (Kyoto University) for sample donation and technical instructions for gene-expression experiments with Thermococcus kodakarensis.
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Fauziah Ma’ruf, I., Sasaki, Y., Kerbs, A. et al. Heterologous gene expression and characterization of two serine hydroxymethyltransferases from Thermoplasma acidophilum. Extremophiles 25, 393–402 (2021). https://doi.org/10.1007/s00792-021-01238-9
- Serine hydroxymethyltransferase
- Threonine aldolase
- Thermoplasma acidophilum