NocO

Enzyme Information
reference: 10.1021/acs.biochem.4c00720
host: E. coli ArcticExpress (DE3)
note: Has improved expression characteristics compared to CylC. They screened 12 phylogenetically diverse CylC homologues and this was the one choosen. Just like in AurF, mutating the metal-binding active site residues causes difficulties with expression. Using E.coli ArcticExpress (DE3), a BL21 derivative solved the issue. In this system Cpn60 and Cpn10 from Oleispira antarctica are constitutively expressed. NocM and NocL copurify with Cpn60 and Cpn10. NocO catalyzes highly chemo-and regioselective chlorination of the terminal carbon of an ACP-tethered dodecanoyl thioester. ,,Mössbauer spectrum of as-isolated NocO exhibits primarily quadrupole doublet features indicative of antiferromagnetically coupled diiron(III) clusters.
producer: Nodularia sp. LEGE 06071
phmm-model: dimetal-carboxylate.hmm

Gene Data
phmm effect mutation gene_name reference
135 catalytic residue Q128 NocO DOI: 10.1021/acs.biochem.4c00720
136 catalytic residue E129 NocO DOI: 10.1021/acs.biochem.4c00720
317 catalytic residue H286 NocO DOI: 10.1021/acs.biochem.4c00720
314 catalytic residue E283 NocO DOI: 10.1021/acs.biochem.4c00720
- loss of chlorination activity H279A NocO DOI: 10.1021/acs.biochem.4c00720
- loss of chlorination activity H132A NocO DOI: 10.1021/acs.biochem.4c00720
- greatly reduced chlorination activity E94A NocO DOI: 10.1021/acs.biochem.4c00720
- loss of chlorination activity E129A NocO DOI: 10.1021/acs.biochem.4c00720
- loss of chlorination activity H286A NocO DOI: 10.1021/acs.biochem.4c00720
- greatly reduced chlorination activity E283A NocO DOI: 10.1021/acs.biochem.4c00720
- greatly reduced chlorination activity; comparable iron stoichometry relative to the wild-type enzyme E252A NocO DOI: 10.1021/acs.biochem.4c00720
- greatly reduced chlorination activity; comparable iron stoichometry relative to the wild-type enzyme Q128A NocO DOI: 10.1021/acs.biochem.4c00720
- greatly reduced chlorination activity; comparable iron stoichometry relative to the wild-type enzyme D282A NocO DOI: 10.1021/acs.biochem.4c00720
313 H-bond acceptors for its proximal iron-binding histidine residues D282 NocO DOI: 10.1021/acs.biochem.4c00720
283 H-bond acceptors for its proximal iron-binding histidine residues E252 NocO DOI: 10.1021/acs.biochem.4c00720
310 catalytic residue H279 NocO DOI: 10.1021/acs.biochem.4c00720
139 catalytic residue H132 NocO DOI: 10.1021/acs.biochem.4c00720
102 catalytic residue E94 NocO DOI: 10.1021/acs.biochem.4c00720