1.14.13.101: senecionine N-oxygenase

This is an abbreviated version!
For detailed information about senecionine N-oxygenase, go to the full flat file.

Reaction

senecionine
+
NADPH
+
H+
+
O2
=
senecionine N-oxide
+
NADP+
+
H2O

Synonyms

GgPNO, oxygenase, senecionine N-, PA N-oxygenase, PNO, senecionine monooxygenase (N-oxide-forming), senecionine N-oxygenase, SNO, TjSNO

ECTree

     1 Oxidoreductases
         1.14 Acting on paired donors, with incorporation or reduction of molecular oxygen
             1.14.13 With NADH or NADPH as one donor, and incorporation of one atom of oxygen into the other donor
                1.14.13.101 senecionine N-oxygenase

Cloned

Cloned on EC 1.14.13.101 - senecionine N-oxygenase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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 4°C
expression in active form in Escherichia coli BL21
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