Table 3.

Assay of in vitro degradation of nPHB granules isolated from R. eutropha^a

Condition	Detected substance	Amt ±SD (mM) by time in min after addition of 1 mM of:
		CoA					3HB-CoA
		5	10	30	60	90	30	90
1. H16 nPHB	CoA	0.84 ± 0.12	0.87 ± 0.06	0.79 ± 0.06	0.68 ± 0.13	0.66 ± 0.06	0.53 ± 0.14	0.49 ± 0.11
	3HB-CoA	0.03 ± 0.03	0.08 ± 0.05	0.14 ± 0.04	0.20 ± 0.06	0.21 ± 0.05	0.45 ± 0.12	0.40 ± 0.13
	Crot-CoA	<0.02	0.04 ± 0.02	0.05 ± 0.02	0.06 ± 0.02	0.06 ± 0.02	0.11 ± 0.05	0.11 ± 0.05
2. H16 ΔphaP1 nPHB	CoA	ND	0.87 ± 0.12	0.88 ± 0.12	0.89 ± 0.10	0.89 ± 0.10	0.60 ± 0.05	0.52 ± 0.11
	3HB-CoA	ND	<0.02	<0.02	0.02 ± 0.02	0.02 ± 0.02	0.50 ± 0.04	0.43 ± 0.02
	Crot-CoA	ND	<0.02	<0.02	<0.02	<0.02	0.16 ± 0.01	0.16 ± 0.01
3. H16 ΔphaP1 ΔphaP2 ΩKm ΔphaP3 ΔphaP4 nPHB	CoA	0.74 ± 0.11	0.69 ± 0.15	0.70 ± 0.09	0.67 ± 0.10	0.63 ± 0.11	0.49 ± 0.03	0.40 ± 0.08
	3HB-CoA	<0.02	<0.02	0.04 ± 0.01	0.05 ± 0.02	0.06 ± 0.02	0.52 ± 0.01	0.43 ± 0.00
	Crot-CoA	<0.02	<0.02	<0.02	0.03 ± 0.02	0.03 ± 0.02	0.17 ± 0.00	0.16 ± 0.00

^aDegradation assays were performed as described in Materials and Methods. The amounts of coenzyme A (CoA), 3-hydroxybutyryl–CoA (3HB-CoA), and crotonyl-CoA (Crot-CoA) were calculated from HPLC peak areas. The values represent the averages from up to 4 assay repetitions and are presented in mM. ND, not determined.