Literature summary extracted from

Dong, Y.; Du, H.; Gao, C.; Ma, T.; Feng, L.
Characterization of two long-chain fatty acid CoA ligases in the Gram-positive bacterium Geobacillus thermodenitrificans NG80-2 (2012), Microbiol. Res., 167, 602-607.

Activating Compound

EC Number	Activating Compound	Comment	Organism	Structure
6.2.1.3	Triton X-100	isoform Facl1 shows 1.3% activity and isoform Facl2 shows 232% activity at 1% (v/v)	Geobacillus thermodenitrificans

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
6.2.1.3	expressed in Escherichia coli BL21(DE3) cells	Geobacillus thermodenitrificans

Inhibitors

EC Number	Inhibitors	Comment	Organism
6.2.1.3	Ag+	isoform Facl1 shows 64% residual activity and isoform Facl2 shows 71% residual activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	Co2+	isoform Facl1 shows 50% residual activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	Cu2+	isoform Facl1 shows 64% residual activity and isoform Facl2 shows 33% residual activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	EDTA	isoform Facl1 shows 12% residual activity and isoform Facl2 shows 26% residual activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	Fe2+	isoform Facl1 shows 57% residual activity and isoform Facl2 shows 36% residual activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	Mn2+	isoform Facl1 shows 28% residual activity and isoform Facl2 shows 40% residual activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	Ni2+	isoform Facl1 shows 1.3% residual activity and isoform Facl2 shows no activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	SDS	isoform Facl1 shows 1.3% residual activity and isoform Facl2 shows 2.6% residual activity at 0.05% (w/v)	Geobacillus thermodenitrificans
6.2.1.3	Zn2+	isoform Facl1 shows 13% residual activity and isoform Facl2 shows 0.36% residual activity at 1 mM	Geobacillus thermodenitrificans

KM Value [mM]

EC Number	KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
6.2.1.3	0.0064	-	caproate	isoform Facl1, at pH 7.5 and 60°C	Geobacillus thermodenitrificans
6.2.1.3	0.032	-	palmitate	isoform Facl2, at pH 7.5 and 60°C	Geobacillus thermodenitrificans

Localization

EC Number	Localization	Comment	Organism	GeneOntology No.	Textmining
6.2.1.3	membrane	-	Geobacillus thermodenitrificans	16020	-

Metals/Ions

EC Number	Metals/Ions	Comment	Organism
6.2.1.3	Ca2+	isoform Facl1 shows 156% activity and isoform Facl2 shows 193% activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	Co2+	isoform Facl2 shows 191% activity at 1 mM	Geobacillus thermodenitrificans
6.2.1.3	additional information	K+, NH4+ and Na+ show no effects on the enzyme activity	Geobacillus thermodenitrificans

Molecular Weight [Da]

EC Number	Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
6.2.1.3	59000	-	2 * 59000, SDS-PAGE	Geobacillus thermodenitrificans
6.2.1.3	118000	-	gel filtration	Geobacillus thermodenitrificans

Organism

EC Number	Organism	UniProt	Comment	Textmining
6.2.1.3	Geobacillus thermodenitrificans	-	-	-
6.2.1.3	Geobacillus thermodenitrificans NG80-2	-	-	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
6.2.1.3	nickel chelating Sepharose column chromatography	Geobacillus thermodenitrificans

Specific Activity [micromol/min/mg]

EC Number	Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
6.2.1.3	2.8	-	isoform Facl1, using caproate as substrate, at pH 7.5 and 60°C	Geobacillus thermodenitrificans
6.2.1.3	8.7	-	isoform Facl2, using palmitate as substrate, at pH 7.5 and 60°C	Geobacillus thermodenitrificans

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
6.2.1.3	ATP + acetate + CoA	isoform Facl1 exhibits less than 10% activity and isoform Facl2 exhibits about 20% activity	Geobacillus thermodenitrificans	AMP + diphosphate + acetyl-CoA	-	?
6.2.1.3	ATP + acetate + CoA	isoform Facl1 exhibits less than 10% activity and isoform Facl2 exhibits about 20% activity	Geobacillus thermodenitrificans NG80-2	AMP + diphosphate + acetyl-CoA	-	?
6.2.1.3	ATP + arachidonate + CoA	isoform Facl1 exhibits about 40% activity and isoform Facl2 exhibits about 60% activity	Geobacillus thermodenitrificans	AMP + diphosphate + arachidonoyl-CoA	-	?
6.2.1.3	ATP + arachidonate + CoA	isoform Facl1 exhibits about 40% activity and isoform Facl2 exhibits about 60% activity	Geobacillus thermodenitrificans NG80-2	AMP + diphosphate + arachidonoyl-CoA	-	?
6.2.1.3	ATP + behenoate + CoA	isoform Facl1 exhibits about 20% activity and isoform Facl2 exhibits about 65% activity	Geobacillus thermodenitrificans	AMP + diphosphate + behenoyl-CoA	-	?
6.2.1.3	ATP + butyrate + CoA	isoform Facl1 exhibits about 20% activity and isoform Facl2 exhibits about 40% activity	Geobacillus thermodenitrificans	AMP + diphosphate + butyryl-CoA	-	?
6.2.1.3	ATP + butyrate + CoA	isoform Facl1 exhibits about 20% activity and isoform Facl2 exhibits about 40% activity	Geobacillus thermodenitrificans NG80-2	AMP + diphosphate + butyryl-CoA	-	?
6.2.1.3	ATP + caproate + CoA	most preferred substrate for isoform Facl1 (100% activity)	Geobacillus thermodenitrificans	AMP + diphosphate + caproyl-CoA	-	?
6.2.1.3	ATP + capryloate + CoA	isoform Facl1 exhibits about 50% activity and isoform Facl2 exhibits about 45% activity	Geobacillus thermodenitrificans	AMP + diphosphate + capryloyl-CoA	-	?
6.2.1.3	ATP + hexacosanoate + CoA	isoform Facl1 exhibits about 8% activity and isoform Facl2 exhibits about 23% activity	Geobacillus thermodenitrificans	AMP + diphosphate + hexacosanoyl-CoA	-	?
6.2.1.3	ATP + laurate + CoA	isoform Facl1 exhibits about 90% activity and isoform Facl2 exhibits about 55% activity	Geobacillus thermodenitrificans	AMP + diphosphate + lauryl-CoA	-	?
6.2.1.3	ATP + lignocerate + CoA	isoform Facl1 exhibits about 20% activity and isoform Facl2 exhibits about 30% activity	Geobacillus thermodenitrificans	AMP + diphosphate + lignoceryl-CoA	-	?
6.2.1.3	ATP + linoleate + CoA	-	Geobacillus thermodenitrificans	AMP + diphosphate + linoleoyl-CoA	-	?
6.2.1.3	ATP + melissoate + CoA	isoform Facl1 exhibits about 3% activity and isoform Facl2 exhibits about 1% activity	Geobacillus thermodenitrificans	AMP + diphosphate + melissoyl-CoA	-	?
6.2.1.3	ATP + myristate + CoA	isoform Facl1 exhibits about 65% activity and isoform Facl2 exhibits about 85% activity	Geobacillus thermodenitrificans	AMP + diphosphate + myristoyl-CoA	-	?
6.2.1.3	ATP + octacosanoate + CoA	isoform Facl1 exhibits about 5% activity and isoform Facl2 exhibits about 3% activity	Geobacillus thermodenitrificans	AMP + diphosphate + octacosanoyl-CoA	-	?
6.2.1.3	ATP + palmitate + CoA	most preferred substrate for isoform Facl2 (100% activity)	Geobacillus thermodenitrificans	AMP + diphosphate + palmitoyl-CoA	-	?
6.2.1.3	ATP + palmitate + CoA	most preferred substrate for isoform Facl2 (100% activity)	Geobacillus thermodenitrificans NG80-2	AMP + diphosphate + palmitoyl-CoA	-	?
6.2.1.3	ATP + stearate + CoA	isoform Facl1 exhibits about 45% activity and isoform Facl2 exhibits about 75% activity	Geobacillus thermodenitrificans	AMP + diphosphate + stearyl-CoA	-	?
6.2.1.3	additional information	unsaturated fatty acids such as 14:1, 16:1, 18:1, 22:1, 18:3(n-3), 20:3(n-6) fatty acids and arachidonic acid are no substrates of isoform Facl1 or Facl2	Geobacillus thermodenitrificans	?	-	?
6.2.1.3	additional information	unsaturated fatty acids such as 14:1, 16:1, 18:1, 22:1, 18:3(n-3), 20:3(n-6) fatty acids and arachidonic acid are no substrates of isoform Facl1 or Facl2	Geobacillus thermodenitrificans NG80-2	?	-	?

Subunits

EC Number	Subunits	Comment	Organism
6.2.1.3	homodimer	2 * 59000, SDS-PAGE	Geobacillus thermodenitrificans

Synonyms

EC Number	Synonyms	Comment	Organism
6.2.1.3	Facl1	isoform	Geobacillus thermodenitrificans
6.2.1.3	Facl2	isoform	Geobacillus thermodenitrificans
6.2.1.3	fatty acid CoA synthetase, AMP forming	-	Geobacillus thermodenitrificans
6.2.1.3	long-chain fatty acid CoA ligase	-	Geobacillus thermodenitrificans

Temperature Optimum [°C]

EC Number	Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
6.2.1.3	60	-	-	Geobacillus thermodenitrificans

Temperature Stability [°C]

EC Number	Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
6.2.1.3	60	-	about 40% of the highest activity of isoform Facl1 still remains after incubation at 60°C for 16 h, and above 50% of the highest isoform Facl2 activity still remains after incubation at 60°C for 8 h	Geobacillus thermodenitrificans

pH Optimum

EC Number	pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.2.1.3	7.5	-	-	Geobacillus thermodenitrificans

pH Stability

EC Number	pH Stability	pH Stability Maximum	Comment	Organism
6.2.1.3	5	11	at pH levels from 5.0 to 11.0, isoform Facl activity increases along with an increase of pH, with the maximum activity obtained at pH 7.5, and about 40% of the highest activity still remains at pH 10.0 for isoform Facl1, and about 40% of the highest activity still remains at pH 9.5 for isoform Facl2	Geobacillus thermodenitrificans

Cofactor

EC Number	Cofactor	Comment	Organism	Structure
6.2.1.3	ATP	-	Geobacillus thermodenitrificans

Expression

EC Number	Organism	Comment	Expression
6.2.1.3	Geobacillus thermodenitrificans	there is a 6.7fold increase and a 10.8fold increase in the mRNA levels of isoforms facl1 and facl2, respectively, when crude oil is used as a sole carbon source instead of sucrose	up

kcat/KM [mM/s]

EC Number	kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
6.2.1.3	9.6	-	palmitate	isoform Facl2, at pH 7.5 and 60°C	Geobacillus thermodenitrificans
6.2.1.3	37.5	-	caproate	isoform Facl1, at pH 7.5 and 60°C	Geobacillus thermodenitrificans