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. 2017 Aug 21;114(36):E7574–E7582. doi: 10.1073/pnas.1619034114

Fig. 3.

Fig. 3.

SerA contributes to 2-KG conversion to d-2-HG. (A) SerA-encoding gene serA is adjacent to the predicated D2HGDH encoding gene d2hgdh. (B) The concentrations of extracellular d-2-HG during growth. (C) The intracellular concentrations of d-2-HG (sampled at 12 h). The strains in B and C were grown in LB medium. Values are the average ± SD (n = 3). (D) The possible configurations of 2-HG produced by SerA-catalyzed 2-KG reduction. (E) HPLC analysis of the product of SerA-catalyzed 2-KG conversion in presence of NADH. The mixtures containing 2-KG (5 mM), NADH (5 mM), native SerA, or heat-inactivated SerA (5.6 μM) in 50 mM Tris (pH 7.4), were incubated at 30 °C for 3 h. (F) The configuration of the SerA-produced 2-HG was determined using an enzymatic method. The reactions using authentic d-2-HG (squares), l-2-HG (circles), or SerA-produced 2-HG (triangles) as substrates were catalyzed by the D2HGDH purified in this study (red lines) or l-2-hydroxyglutarate oxidoreductase (LGRO, blue lines), respectively. The purification and characterization of LGRO are presented in SI Appendix, Fig. S4 F and G.