4FM4

Wild Type Fe-type Nitrile Hydratase from Comamonas testosteroni Ni1

Summary for 4FM4

• Entry DOI: 10.2210/pdb4fm4/pdb
• Descriptor: Nitrile hydratase alpha subunit, Nitrile hydratase beta subunit, PHOSPHATE ION, ... (5 entities in total)
• Functional Keywords: iron type hydratase, hydrolysis, sulfinic acid, lyase
• Biological source: Comamonas testosteroni; More
• Total number of polymer chains: 16
• Total formula weight: 368152.31

Authors

Kuhn, M.L.,Martinez, S.,Gumataotao, N.,Bornscheuer, U.,Liu, D.,Holz, R.C. (deposition date: 2012-06-15, release date: 2012-08-22, Last modification date: 2024-11-27)

Primary citation

Kuhn, M.L.,Martinez, S.,Gumataotao, N.,Bornscheuer, U.,Liu, D.,Holz, R.C.
The Fe-type nitrile hydratase from Comamonas testosteroni Ni1 does not require an activator accessory protein for expression in Escherichia coli.
Biochem.Biophys.Res.Commun., 424:365-370, 2012

PubMed Abstract: We report herein the functional expression of an Fe-type nitrile hydratase (NHase) without the co-expression of an activator protein or the Escherichia coli chaperone proteins GroES/EL. Soluble protein was obtained when the α- and β-subunit genes of the Fe-type NHase Comamonas testosteroni Ni1 (CtNHase) were synthesized with optimized E. coli codon usage and co-expressed. As a control, the Fe-type NHase from Rhodococcus equi TG328-2 (ReNHase) was expressed with (ReNHase(+Act)) and without (ReNHase(-Act)) its activator protein, establishing that expression of a fully functional, metallated ReNHase enzyme requires the co-expression of its activator protein, similar to all other Fe-type NHase enzymes reported to date, whereas the CtNHase does not. The X-ray crystal structure of CtNHase was determined to 2.4Å resolution revealing an αβ heterodimer, similar to other Fe-type NHase enzymes, except for two important differences. First, two His residues reside in the CtNHase active site that are not observed in other Fe-type NHase enzymes and second, the active site Fe(III) ion resides at the bottom of a wide solvent exposed channel. The solvent exposed active site, along with the two active site histidine residues, are hypothesized to play a role in iron incorporation in the absence of an activator protein.

PubMed: 22713452
DOI: 10.1016/j.bbrc.2012.06.036

Experimental method

X-RAY DIFFRACTION (2.384 Å)