Part:BBa_K4011003
Fre-SttH
Fre-SttH is a fused protein used to halogenate the 6th carbon of the tryptophan in the dye production. Fre-SttH is composed of two separate domains, fre and sttH. Fre is from E.coli and SttH is from Streptomyces toxytricini. They are fused by a rigid linker with the protein sequence EAAAKEAAAK. SttH is a trp-6-haloganese that requires FADH2 as a cofactor to convert trp into 6-X-trp, and is highly insoluble in E. coli. Therefore, Fre, a highly-soluble flavin reductase which reduces FAD to FADH2 from E. coli, is fused with SttH as a N-terminal soluble tag, enabling the protein to become soluble and eliminating the need for costly FADH2 cofactors to be added. The fre-sttH in ptac system helps to improve the solubility of it and also allows it to be expressed in any strain of E.coli. The activity and substrate-product concentrations of fre-sttH is measured using HPLC.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 576
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 948
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 424
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