22.214.171.124: senecionine N-oxygenase
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For detailed information about senecionine N-oxygenase, go to the full flat file.
flavin-dependent monooxygenase, fmoa, GgPNO, oxygenase, senecionine N-, PA N-oxygenase, PNO, pyrrolizidine alkaloid N-oxygenase, senecionine monooxygenase (N-oxide-forming), senecionine N-oxygenase, SNO, TjSNO, ZvFMOa, ZvPNO
1.14.13 With NADH or NADPH as one donor, and incorporation of one atom of oxygen into the other donor
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Cloned on EC 126.96.36.199 - senecionine N-oxygenase
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expressed in Escherichia coli BL21(DE3) cells
expressing GgPNO without the signal peptide in at least a partially soluble and active form. Betaine is added to the medium and promotes its uptake by osmotic stress by sorbitol. Further improvements are achieved by coexpression with the Escherichia coli chaperones DnaK/DnaJ/GrpE by using the plasmid pRDKJG and by reducing the expression temperature to 4Â°C
expression in active form in Escherichia coli BL21
gene fmoa, recombinant expression of His6-tagged wild-type and mutant isozymes ZvPNO in Escherichia coli strain BL21(DE3)
gene pno, recombinant expression of His6-tagged isozyme ZvPNO in Escherichia coli strain BL21(DE3)
the heterologous expression results in the formation of an insoluble and inactive protein. Despite the finding that protein solubilization and subsequent renaturation results in an active protein, the specific activity remains low. Active SNO is detected by using an an Sf9 insect cell expression system, but the yield of protein is too low for further biochemical characterization. Using an alignment of senecionine N-oxygenase of Tyria jacobaeae and of flavin-dependent monooxygenase-like sequences of the dipteran species Drosophila melanogaster and Anopheles gambiae, several new flavin-dependent monooxygenase sequences of arctiids and other Lepidoptera are idenified