TABLE 3.
Reaction requirements for isobutene formation by purified His6-ScMDD and His6-ScMDD2a
| Assay component | Amt of isobutene (pmol min−1 mg protein−1) produced by: |
|
|---|---|---|
| His6-ScMDD | His6-ScMDD2 | |
| Complete | 1.33 ± 0.27 | 6.44 ± 0.95 |
| −ATP | ND | ND |
| −3-HMB | ND | ND |
| Boiled enzyme | ND | ND |
a
The complete assay contained 500 μg ScMDD, 333 mM 3-HMB, 250 mM ATP, and standard assay components. No isobutene was detected when ATP or 3-HMB was omitted (−ATP and −3-HMB, respectively) from assay mixtures or when boiled enzyme was used in place of purified ScMDD. ND, none detected.