Literature summary for 2.1.1.140 extracted from

Bennett, M.R.; Thompson, M.L.; Shepherd, S.A.; Dunstan, M.S.; Herbert, A.J.; Smith, D.R.M.; Cronin, V.A.; Menon, B.R.K.; Levy, C.; Micklefield, J.
Structure and biocatalytic scope of coclaurine N-methyltransferase (2018), Angew. Chem. Int. Ed. Engl., 57, 10600-10604 .

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Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Coptis japonica

Crystallization (Commentary)

Crystallization (Comment)	Organism
in complex with S-adenosyl-L-homocysteine and N-methylheliamine. Molecular modeling of structure with natural substrate coclaurine.Both enantiomers of coclaurine can be accommodated in the CNMT active site	Coptis japonica

Protein Variants

Protein Variants	Comment	Organism
E204A	4-6% of wild-type activity	Coptis japonica
E207A	60-75% of wild-type activity	Coptis japonica
F332A	0.6% of wild-typ activity with substrate coclaurine	Coptis japonica
G331A	negligible effect on activity	Coptis japonica
H208A	1% of wild-type activity with substrate heliamine, 4% with substrate coclaurine, respectively	Coptis japonica
R330A	negligible effect on activity	Coptis japonica
W329A	4% of wild-type activity with substrate heliamine, 0.5% with substrate coclaurine, respectively	Coptis japonica
Y328A	5% of wild-type activity with substrate heliamine, 10% with substrate coclaurine, respectively	Coptis japonica

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.265	-	(RS)-coclaurine	wild-type, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.311	-	heliamine	wild-type, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.601	-	(RS)-coclaurine	mutant Y328A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
1.033	-	(RS)-coclaurine	mutant W329A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
1.565	-	heliamine	mutant W329A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
1.726	-	(RS)-coclaurine	mutant F332A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
2.096	-	heliamine	mutant Y328A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica

Organism

Organism	UniProt	Comment	Textmining
Coptis japonica	Q948P7	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
S-adenosyl-L-methionine + (RS)-coclaurine	-	Coptis japonica	S-adenosyl-L-homocysteine + (RS)-N-methylcoclaurine	-	?
S-adenosyl-L-methionine + heliamine	-	Coptis japonica	S-adenosyl-L-homocysteine + N-methylheliamine	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.01	-	(RS)-coclaurine	mutant W329A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.02	-	(RS)-coclaurine	mutant F332A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.12	-	(RS)-coclaurine	mutant Y328A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.14	-	heliamine	mutant W329A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.2	-	heliamine	mutant Y328A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.53	-	(RS)-coclaurine	wild-type, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.6	-	heliamine	wild-type, pH not specified in the publication, temperature not specified in the publication	Coptis japonica

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.01	-	(RS)-coclaurine	mutant F332A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.01	-	(RS)-coclaurine	mutant W329A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.09	-	heliamine	mutant W329A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.09	-	heliamine	mutant Y328A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
0.2	-	(RS)-coclaurine	mutant Y328A, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
2	-	(RS)-coclaurine	wild-type, pH not specified in the publication, temperature not specified in the publication	Coptis japonica
2	-	heliamine	wild-type, pH not specified in the publication, temperature not specified in the publication	Coptis japonica

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Release 2023.1 (January 2023)