Table 1. Enzymes used in the synthesis of DPM.
aEnzyme | EC # | Gene | Source | Substrate | d K m (mM) | d k cat (sec−1) |
bACS | 6.2.1.1 | Acs1 | S. cerevisiae | Acetate CoA | 0.28 0.24 | 10 |
cACT | 2.3.1.9 | mvaE | E. faecalis | Ac-CoA | 0.60 | 2.3 |
cHMGS2 | 2.3.3.10 | mvaS | E. faecalis | acac-CoA Ac-CoA | 0.015 0.35 | 1.0 |
cThRed | 1.1.1.34 | mvaE | E. faecalis | HMG-CoA | 0.023 | 0.55 |
cHMGR | 1.1.1.34 | mvaE | E. faecalis | HMG-CoA | 0.020 | 0.67 |
cMVK | 2.7.1.36 | mvaK1 | S. aureus | Mev | 0.027 | 19 |
cPMK | 2.7.4.2 | mvaK2 | S. pneumoniae | P-mev | 0.0042 | 5.0 |
bPK | 2.7.1.40 | PKM2 | O. cuniculus | PEP | 0.040 | 160 |
bMK | 2.7.4.3 | AK1 | O. cuniculus | AMP | e0.50 | e410 |
bPPiase | 3.6.1.1 | Ppa1 | S. cerevisiae | PPi | e0.0050 | e260 |
Abbreviations: ACS, acetyl-CoA synthetase; ACT, acetoacetyl-CoA thiolase; HMGS2, HMG-CoA synthase; ThRed, acetoacetyl-CoA thiolase/HMG-CoA reductase (dual-function enzyme); HMGR, HMG-CoA reductase; MVK, mevalonate kinase; PMK, phosphomevalonate kinase; PK, pyruvate kinase; MK, myokinase; PPiase, inorganic pyrophosphatase.
Obtained from commercial sources.
Expressed in E. coli and purified.
Standard errors are <5% in all cases (see Materials and Methods ).