Part:BBa_K1137001
M. Smegmatis FprA
This biobrick encodes a NADPH-ferredoxin reductase FprA. This replenishes FdxA by reducing it with NADPH as an electron donor and is required for the proper function of BBa_K1137000 along with BBa_K1137001 in the following reaction:
(EC=1.18.1.2)
NADPH+ 〖2FdxA〗_ox ↔ 〖NADP〗^++ 〖2FdxA〗_red+ H^+
FprA in addition to SirA has low homology to mammalian proteins and contains one tightly bound FAD. Additonally it has been well characterized and it’s kinetics determined experimentally. As mentioned under BBa_K1137000, even with minimal induction in a duet vector expression system, we were able to obtain growth in M9 media supplemented with glucose as a carbon source when co-expressed with BBa_K1137001 and BBa_K1137002, however the parent strain which lacked these biobricks was unable to grow. FprA from M. smegmatis has a homolog in M. tuberculosis.
Growth curves: BL21 (DE3) ΔcysI containing the MycoSIR pathway (MycoSIR E.coli) were grown in liquid minimal media containing Various concentration of IPTG. (A) Replicates of each strain were measured for absorbance in a spectrophotometer every 10 minutes for 14 hours. Growth was observed for the WT BL21 E.coli, (blue), and the MycoSIR E.coli (red). No growth was detected for uninduced MycoSIR E.coli (purple) or for the BL21 (DE3) ΔcysI that did not contain the synthetic pathway (Orange) . (B) Mean Final ODs of all replicates, measured after 14 hours of growth. Growth was detected in zmSIR E.coli and WT BL21 but not in uninduced zmSIR strain.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 948
Illegal AgeI site found at 310
Illegal AgeI site found at 928
Illegal AgeI site found at 1252
Illegal AgeI site found at 1306 - 1000COMPATIBLE WITH RFC[1000]
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