Literature summary for 3.5.1.91 extracted from

Hashimoto, Y.; Sakashita, T.; Fukatsu, H.; Sato, H.; Kobayashi, M.
A new synthetic route to N-benzyl carboxamides through the reverse reaction of N-substituted formamide deformylase (2014), Appl. Environ. Microbiol., 80, 61-69.

Activating Compound

Activating Compound	Comment	Organism
ammonium persulfate	12% activation at 1 mM	Arthrobacter pascens
diisopropyl fluorophosphate	17% activation at 1 mM	Arthrobacter pascens
EDTA	27% activation at 1 mM	Arthrobacter pascens
KCN	slight activation at 1 mM	Arthrobacter pascens

Inhibitors

Inhibitors	Comment	Organism
2,2'-dipyridyl	15% inhibition at 1 mM	Arthrobacter pascens
2-mercaptoethanol	64% inhibition at 1 mM	Arthrobacter pascens
3-Phenylpropylamine	inhibits the reverse enzyme reaction	Arthrobacter pascens
8-hydroxyquinoline	84% inhibition at 1 mM	Arthrobacter pascens
AgNO3	complete inhibition at 1 mM	Arthrobacter pascens
aminoguanidine	19% inhibition at 1 mM	Arthrobacter pascens
aniline	a benzylamine analogue, dead-end inhibition study, overview; inhibits the reverse enzyme reaction, uncompetitive versus benzylamine, competitive versus formate	Arthrobacter pascens
Butyrate	slight inhibition of the reverse reaction, competitive versus benzylamine, noncompetitive versus formate	Arthrobacter pascens
CdCl2	57% inhibition at 1 mM	Arthrobacter pascens
CuCl	complete inhibition at 0.25 mM	Arthrobacter pascens
CuCl2	complete inhibition at 1 mM	Arthrobacter pascens
dithiothreitol	complete inhibition at 1 mM	Arthrobacter pascens
H2O2	slight inhibition at 1 mM	Arthrobacter pascens
HgCl2	complete inhibition at 1 mM	Arthrobacter pascens
Isobutyrate	slight inhibition of the reverse reaction	Arthrobacter pascens
additional information	no or poor inhibition by 1 mM of LiCl, NaCl, MgCl2, CaCl2, BaCl2, MnCl2, AlCl3, Pb(NO3)2, FeSO4, FeCl3, RbCl, SrCl2, CsCl, Na2MoO4, CoCl2, NiCl2, and ZnCl2. No inhibition by iodoacetate, 5,5'-dithio-bis-2-nitrobenzoate, diethyldithiocarbamate, NaN3, and phenylmethanesulfonyl fluoride. Valerate, isovalerate, and caproate cannot be tested because they are insoluble in the reaction buffer	Arthrobacter pascens
N-ethylmaleimide	79% inhibition at 1 mM	Arthrobacter pascens
o-phenanthroline	32% inhibition at 1 mM	Arthrobacter pascens
p-chloromercuribenzoate	complete inhibition at 1 mM	Arthrobacter pascens
phenethylamine	inhibits the reverse enzyme reaction	Arthrobacter pascens
phenylhydrazine	23% inhibition at 1 mM	Arthrobacter pascens
Semicarbazide	8% inhibition at 1 mM	Arthrobacter pascens
SnCl2	44% inhibition at 1 mM	Arthrobacter pascens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	bisubstrate kinetic analysis using formate and benzylamine, overview	Arthrobacter pascens
0.075	-	N-benzylformamide	pH 7.5, 25°C	Arthrobacter pascens
53.6	-	benzylamine	pH 7.0, 25°C	Arthrobacter pascens
1210	-	acetate	pH 7.0, 25°C	Arthrobacter pascens
3010	-	formate	pH 7.0, 25°C	Arthrobacter pascens
3850	-	propionate	pH 7.0, 25°C	Arthrobacter pascens

Metals/Ions

Metals/Ions	Comment	Organism	Structure
additional information	no or poor inhibition by 1 mM of LiCl, NaCl, MgCl2, CaCl2, BaCl2, MnCl2, AlCl3, Pb(NO3)2, FeSO4, FeCl3, RbCl, SrCl2, CsCl, Na2MoO4, CoCl2, NiCl2, and ZnCl2	Arthrobacter pascens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
N-benzylformamide + H2O	Arthrobacter pascens	-	formate + benzylamine	-	r
N-benzylformamide + H2O	Arthrobacter pascens F164	-	formate + benzylamine	-	r

Organism

Organism	UniProt	Comment	Textmining
Arthrobacter pascens	-	-	-
Arthrobacter pascens F164	-	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
N-benzylformamide + H2O = formate + benzylamine	the reverse reaction of this enzyme proceeds via an ordered two-substrate, two-product (bi-bi) mechanism in which formate binds first to the enzyme active site, followed by benzylamine binding and the subsequent release of N-benzylformamide, kinetic mechanism, overview	Arthrobacter pascens	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	mass spectra on LC-ESI-MS of the reverse-reaction products of N-substituted formamide deformylase, overview. No activity with butyrate and isobutyrate	Arthrobacter pascens	?	-	?
additional information	mass spectra on LC-ESI-MS of the reverse-reaction products of N-substituted formamide deformylase, overview. No activity with butyrate and isobutyrate	Arthrobacter pascens F164	?	-	?
N-benzylacetamide + H2O	the enzyme catalyzes the reverse reaction from acetate and benzylamine at high substrate concentrations	Arthrobacter pascens	acetate + benzylamine	-	r
N-benzylacetamide + H2O	the enzyme catalyzes the reverse reaction from acetate and benzylamine at high substrate concentrations	Arthrobacter pascens F164	acetate + benzylamine	-	r
N-benzylformamide + H2O	-	Arthrobacter pascens	formate + benzylamine	-	r
N-benzylformamide + H2O	the enzyme catalyzes the reverse reaction at high substrate concentrations, synthesizing N-benzylformamide from benzylamine and formate. The reverse reaction proceeds only in the presence of high substrate concentrations. Effects of pH and inhibitors on the reverse reaction are almost the same as those on the forward reaction, suggesting that the forward and reverse reactions are both catalyzed at the same catalytic site	Arthrobacter pascens	formate + benzylamine	-	r
N-benzylformamide + H2O	-	Arthrobacter pascens F164	formate + benzylamine	-	r
N-benzylformamide + H2O	the enzyme catalyzes the reverse reaction at high substrate concentrations, synthesizing N-benzylformamide from benzylamine and formate. The reverse reaction proceeds only in the presence of high substrate concentrations. Effects of pH and inhibitors on the reverse reaction are almost the same as those on the forward reaction, suggesting that the forward and reverse reactions are both catalyzed at the same catalytic site	Arthrobacter pascens F164	formate + benzylamine	-	r
N-benzylpropionamide + H2O	the enzyme catalyzes the reverse reaction from propionate and benzylamine at high substrate concentrations	Arthrobacter pascens	propionate + benzylamine	-	r
N-benzylpropionamide + H2O	the enzyme catalyzes the reverse reaction from propionate and benzylamine at high substrate concentrations	Arthrobacter pascens F164	propionate + benzylamine	-	r

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	reverse reaction	Arthrobacter pascens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
45	-	pH 7.5, 30 min, completely stable up to	Arthrobacter pascens
50	-	pH 7.5, 30 min, 78% activity remaining	Arthrobacter pascens
55	-	pH 7.5, 30 min, and above, no activity remaining	Arthrobacter pascens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	reverse reaction	Arthrobacter pascens

pH Range

pH Minimum	pH Maximum	Comment	Organism
5	9	activity range, profile overview	Arthrobacter pascens

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
6.5	9.5	25°C, 30 min, 80% activity remaining at pH 8.0, below 20% at pH 6,5 and pH 9.5, inactivation at pH 6.0 and pH 10.0, profile overview	Arthrobacter pascens

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
13.3	-	pH 7.0, 25°C	Arthrobacter pascens	3-Phenylpropylamine
14.5	-	pH 7.0, 25°C	Arthrobacter pascens	aniline
27.8	-	pH 7.0, 25°C	Arthrobacter pascens	phenethylamine
275	-	pH 7.0, 25°C	Arthrobacter pascens	Butyrate
280	-	pH 7.0, 25°C	Arthrobacter pascens	Isobutyrate

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the the amidohydrolase superfamily. Although N-substituted formamide deformylase is one of the amine-forming deformylases, it shows no amino acid sequence similarity to any other deformylases known so far. Contrary to expectation, the deduced amino acid sequence of N-substituted formamide deformylase exhibited the highest overall sequence identity (28%) to those of regulatory proteins	Arthrobacter pascens
additional information	the enzyme contains (alpha/beta)8 barrel structure in the N-terminal region with five conserved residues (four histidines and one aspartic acid)	Arthrobacter pascens
physiological function	N-substituted formamide deformylase is involved in the degradation of toxic isonitriles	Arthrobacter pascens

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.003	-	propionate	pH 7.0, 25°C	Arthrobacter pascens
0.008	-	acetate	pH 7.0, 25°C	Arthrobacter pascens
0.011	-	formate	pH 7.0, 25°C	Arthrobacter pascens
0.411	-	benzylamine	pH 7.0, 25°C	Arthrobacter pascens
687	-	N-benzylformamide	pH 7.5, 25°C	Arthrobacter pascens