Combinatorial optimization of synthetic operons for the microbial production of p-coumaryl alcohol with Escherichia coli

Table 1 81 Synthetic p-coumaryl alcohol operon variants differing in the combination of the SD sequence–START codon spacings (5, 9, 13 nt)

Variant	SD seq.–START spacing	Variant	SD seq.–START spacing	Variant	SD seq.–START spacing
1	5-5-5-5	28	9-5-5-5	55	13-5-5-5
2	5-5-5-9	29	9-5-5-9	56	13-5-5-9
3	5-5-5-13	30	9-5-5-13	57	13-5-5-13
4	5-5-9-5	31	9-5-9-5	58	13-5-9-5
5	5-5-9-9	32	9-5-9-9	59	13-5-9-9
6	5-5-9-13	33	9-5-9-13	60	13-5-9-13
7	5-5-13-5	34	9-5-13-5	61	13-5-13-5
8	5-5-13-9	35	9-5-13-9	62	13-5-13-9
9	5-5-13-13	36	9-5-13-13	63	13-5-13-13
10	5-9-5-5	37	9-9-5-5	64	13-9-5-5
11	5-9-5-9	38	9-9-5-9	65	13-9-5-9
12	5-9-5-13	39	9-9-5-13	66	13-9-5-13
13	5-9-9-5	40	9-9-9-5	67	13-9-9-5
14	5-9-9-9	41	9-9-9-9	68	13-9-9-9
15	5-9-9-13	42	9-9-9-13	69	13-9-9-13
16	5-9-13-5	43	9-9-13-5	70	13-9-13-5
17	5-9-13-9	44	9-9-13-9	71	13-9-13-9
18	5-9-13-13	45	9-9-13-13	72	13-9-13-13
19	5-13-5-5	46	9-13-5-5	73	13-13-5-5
20	5-13-5-9	47	9-13-5-9	74	13-13-5-9
21	5-13-5-13	48	9-13-5-13	75	13-13-5-13
22	5-13-9-5	49	9-13-9-5	76	13-13-9-5
23	5-13-9-9	50	9-13-9-9	77	13-13-9-9
24	5-13-9-13	51	9-13-9-13	78	13-13-9-13
25	5-13-13-5	52	9-13-13-5	79	13-13-13-5
26	5-13-13-9	53	9-13-13-9	80	13-13-13-9
27	5-13-13-13	54	9-13-13-13	81	13-13-13-13

All variants of the operon were cloned into the pALXtreme vector backbone [16] (general scheme: pALXtreme-1a_T7_prom>x-Rstal-x-Pc4cl-x-Zmccr-x-Zmcad) and subsequently transformed to E. coli BL21 (DE3) lacI ^Q1 [relevant characteristics of the genotype: F⁻ ompT hsdSB(rB⁻, mB⁻) gal dcm (DE3)].

ISSN: 1475-2859