Literature summary for 6.2.1.12 extracted from

Stuible, H.; Buttner, D.; Ehlting, J.; Hahlbrock, K.; Kombrink, E.
Mutational analysis of 4-coumarate:CoA ligase identifies functionally important amino acids and verifies its close relationship to other adenylate-forming enzymes (2000), FEBS Lett., 467, 117-122.

Search for more literature for 6.2.1.12

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli M15[pRP4] as His-tag fusion protein	Arabidopsis thaliana

Protein Variants

Protein Variants	Comment	Organism
C403A	activity reduced to 45% of wild-type level, no effect on Km values for ATP and caffeate	Arabidopsis thaliana
E401Q	activity reduced to 21% of wild-type level, no effect on Km values for ATP and caffeate	Arabidopsis thaliana
K211S	activity reduced to 2.7% of wild-type level	Arabidopsis thaliana
K445T	activity reduced to 0.2% of wild-type level	Arabidopsis thaliana
K457S	activity reduced to 4% of wild-type level	Arabidopsis thaliana
K540N	no activity	Arabidopsis thaliana
R449Q	activity reduced to 3% of wild-type level, Km values significantly increased	Arabidopsis thaliana

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.005	-	caffeate	K457S mutant enzyme	Arabidopsis thaliana
0.014	-	caffeate	K211S mutant enzyme	Arabidopsis thaliana
0.018	-	caffeate	C403A mutant enzyme	Arabidopsis thaliana
0.024	-	caffeate	wild type	Arabidopsis thaliana
0.028	-	caffeate	E401Q mutant enzyme	Arabidopsis thaliana
0.029	-	ATP	K457S mutant enzyme	Arabidopsis thaliana
0.151	-	ATP	K211S mutant enzyme	Arabidopsis thaliana
0.163	-	ATP	wild type	Arabidopsis thaliana
0.173	-	ATP	C403 mutant enzyme	Arabidopsis thaliana
0.24	-	ATP	E401Q mutant enzyme	Arabidopsis thaliana
0.4	-	caffeate	R449Q mutant enzyme	Arabidopsis thaliana
1.507	-	ATP	R449Q mutant enzyme	Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + 4-coumarate + CoA	Arabidopsis thaliana	plant phenylpropanoid metabolism	AMP + diphosphate + 4-coumaroyl-CoA	-	?

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	-	isoform 4CL2	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme using His-tag	Arabidopsis thaliana

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.34	-	K211S mutant enzyme with caffeate as substrate	Arabidopsis thaliana
0.37	-	R449Q mutant enzyme with caffeate as substrate	Arabidopsis thaliana
0.46	-	K457S mutant enzyme with caffeate as substrate	Arabidopsis thaliana
2.64	-	E401Q mutant enzyme with caffeate as substrate	Arabidopsis thaliana
5.64	-	C403A mutant enzyme with caffeate as substrate	Arabidopsis thaliana
12.5	-	wild type with caffeate as substrate	Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + 4-coumarate + CoA	-	Arabidopsis thaliana	AMP + diphosphate + 4-coumaroyl-CoA	-	?
ATP + 4-coumarate + CoA	plant phenylpropanoid metabolism	Arabidopsis thaliana	AMP + diphosphate + 4-coumaroyl-CoA	-	?
ATP + caffeate + CoA	-	Arabidopsis thaliana	AMP + diphosphate + caffeoyl-CoA	-	?

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Release 2023.1 (January 2023)