TABLE 2.

Porphyrin accumulation in E. coli transformants overexpressing different combinations of heme biosynthetic genes^a

Combination	Heme biosynthetic genes overexpressed on:			Concn (μmol/liter)b
	pACmod	pBBR1-MCS2	pUCmod	Uropophyrin I	Uroporphyrin III	Pentacarboxy-porphyrin	Coproporphyrin I	Coproporphyrin III	Protoporphyrin	Heme	Total
1				NDc	ND	ND	ND	ND	Trd	0.2 ± 0.0	0.2 ± 0.0
2	A			3.3 ± 0.8	0.6 ± 0.2	ND	0.3 ± 0.0	2.0 ± 0.1	0.6 ± 0.5	0.9 ± 0.1	7.7 ± 1.6
3	A, B			4.0 ± 1.8	0.6 ± 0.3	ND	0.1 ± 0.0	0.9 ± 0.2	0.6 ± 0.1	0.4 ± 0.1	6.7 ± 2.4
4	A, B, D			0.8 ± 0.1	1.8 ± 0.1	ND	0.1 ± 0.0	1.0 ± 0.3	1.1 ± 0.1	0.5 ± 0.1	5.3 ± 0.7
5	A, B, C			14.4 ± 0.2	ND	1.1 ± 0.2	0.5 ± 0.1	ND	Tr	0.2 ± 0.1	16.2 ± 0.5
6	A, B, C, D			7.2 ± 1.8	41 ± 10	ND	ND	1.6 ± 0.3	0.2 ± 0.1	0.4 ± 0.3	49 ± 12
7	A, B, C	E		8.2 ± 0.1	ND	2.2 ± 0.1	1.0 ± 0.1	0.2 ± 0.0	Tr	0.2 ± 0.1	11.7 ± 0.4
8	A, B, C, D	E		ND	ND	ND	3.0 ± 0.2	47 ± 4	0.4 ± 0.2	0.4 ± 0.1	51 ± 4
9	A, B, C, D	E, F		ND	ND	ND	0.3 ± 0.1	0.7 ± 0.2	82 ± 6	2.4 ± 0.5	85 ± 7
10	A, B, C, D	E, F	H	ND	ND	ND	0.3 ± 0.1	1.0 ± 0.2	84 ± 8	3.3 ± 0.3	89 ± 9
11	A, B, C, D		Y	2.0 ± 0.2	7.2 ± 0.6	ND	ND	1.4 ± 0.3	Tr	0.1 ± 0.1	11 ± 1
12	A, B, C, D	F	Y	0.7 ± 0.1	4.8 ± 0.1	ND	ND	2.6 ± 0.1	0.5 ± 0.1	0.3 ± 0.1	9.3 ± 0.4
13	A, B, C, D	E	Y	1.3 ± 0.3	0.2 ± 0.1	ND	ND	37 ± 2	Tr	0.7 ± 0.0	39 ± 2
14	A, B, C	E	Y	7.0 ± 0.7	ND	4.7 ± 0.6	2.1 ± 0.4	1.8 ± 0.3	ND	0.2 ± 0.1	16 ± 2
15	A, B, C, D	E, F	Y	0.7 ± 0.3	0.4 ± 0.1	ND	1.5 ± 0.2	21 ± 3	28 ± 4	1.7 ± 0.4	52 ± 8

^aE. coli cells were cultured in LB media containing chloramphenicol, kanamycin, and carbenicillin for 48 h at 30°C.

^bThe values are means ± standard deviations for three experiments.

^cND, not detected.

^dTr, trace (less than 0.1 μmol/liter).