TABLE 2.
In vitro reconstitution of the serine degradation pathway by using recombinant purified T. maritima enzymes
| Substrate(s) added | Enzyme(s) added | Conversion of NADH to NAD+ and/or ATP to ADP |
|---|---|---|
| l-Serine and pyruvatea | SAT and HPR | + |
| SAT | − | |
| HPR | − | |
| 3-Hydroxypyruvateb | HPR and GK-II | + |
| HPR | − | |
| GK-II | − |
a
The conversion of l-serine to d-glycerate by a mixture of the enzymes SAT (TM1400) and HPR (TM1401) was determined by monitoring the decrease in absorbance at 340 nm due to the conversion of NADH to NAD+ (see the assay shown in Fig. 3B).
b
The conversion of hydroxypyruvate to 2-phospho-d-glycerate by a mixture of HPR (TM1400) and GK-II (TM1585) was detected by monitoring the conversion of NADH to NAD+ and ATP to ADP, using HPLC.