| Autor: |
Samsonov VV; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia., Kuznetsova AA; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia., Rostova JG; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia., Samsonova SA; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia., Ziyatdinov MK; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia., Kiriukhin MY; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia. |
| Jazyk: |
angličtina |
| Zdroj: |
Microorganisms [Microorganisms] 2022 Aug 31; Vol. 10 (9). Date of Electronic Publication: 2022 Aug 31. |
| DOI: |
10.3390/microorganisms10091766 |
| Abstrakt: |
In E. coli and P. ananatis , L-serine biosynthesis is initiated by the action of D-3-phosphoglycerate dehydrogenase (SerA), which converts D-3-phosphoglycerate into 3-phosphohydroxypyruvate. SerA can concomitantly catalyze the production of D-2-hydroxyglutarate (D-2-HGA) from 2-ketoglutarate by oxidizing NADH to NAD + . Several bacterial D-2-hydroxyglutarate dehydrogenases (D2HGDHs) have recently been identified, which convert D-2-HGA back to 2-ketoglutarate. However, knowledge about the enzymes that can metabolize D-2-HGA is lacking in bacteria belonging to the Enterobacteriaceae family. We found that ydiJ encodes novel D2HGDHs in P. ananatis and E. coli , which were assigned as D2HGDH Pa and D2HGDH Ec , respectively. Inactivation of ydiJ in P. ananatis and E. coli led to the significant accumulation of D-2-HGA. Recombinant D2HGDH Ec and D2HGDH Pa were purified to homogeneity and characterized. D2HGDH Ec and D2HGDH Pa are homotetrameric with a subunit molecular mass of 110 kDa. The pH optimum was 7.5 for D2HGDH Pa and 8.0 for D2HGDH Ec . The Km for D-2-HGA was 208 μM for D2HGDHPa and 83 μM for D2HGDH Ec . The enzymes have strict substrate specificity towards D-2-HGA and displayed maximal activity at 45 °C. Their activity was completely inhibited by 0.5 mM Mn 2+ , Ni 2+ or Co 2+ . The discovery of a novel family of D2HGDHs may provide fundamental information for the metabolic engineering of microbial chassis with desired properties. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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