Literature summary for 4.1.2.13 extracted from

Labbe, G.; de Groot, S.; Rasmusson, T.; Milojevic, G.; Dmitrienko, G.I.; Guillemette, J.G.
Evaluation of four microbial Class II fructose 1,6-bisphosphate aldolase enzymes for use as biocatalysts (2011), Protein Expr. Purif., 80, 224-233.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Pseudomonas aeruginosa
expressed in Escherichia coli BL21(DE3) cells	Bacillus cereus
expressed in Escherichia coli BL21(DE3) cells	Mycobacterium tuberculosis
expressed in Escherichia coli BL21(DE3) cells	Pyricularia grisea

Inhibitors

Inhibitors	Comment	Organism
EDTA	the specific activity of the Bacillus cereus aldolase is partially restored after EDTA inactivation by Co2+ and Cd2+, as well as Zn2+, but not significantly restored by Cu2+, Mn2+, Mg2+, or Ni2+	Bacillus cereus
EDTA	the activity of the EDTA-inactivated enzyme increases more than 6fold upon the addition of 0.002 mM Zn2+ and increases by a factor 2 with the addition of 0.002 mM Co2+. The addition of Cu2+, Ni2+, Cd2+, Mn2+, or Mg2+ does not reactivate the enzyme significantly	Mycobacterium tuberculosis
EDTA	the specific activity of the recombinant Pseudomonas aeruginosa aldolase is 5.5times higher after inactivation by EDTA followed by the addition of 0.02 mM cobalt chloride, and increases more than 2fold after EDTA inactivation followed by the addition of 0.1 mM manganese(II)-chloride	Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0279	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Mycobacterium tuberculosis
0.035	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Pseudomonas aeruginosa
0.051	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Pyricularia grisea
0.45	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Bacillus cereus

Metals/Ions

Metals/Ions	Comment	Organism
additional information	the addition of Co2+ to the purified Bacillus cereus aldolase does not increase the relatively low specific activity of this recombinant enzyme	Bacillus cereus
Zn2+	contains 0.16 zinc atoms per subunit	Pseudomonas aeruginosa
Zn2+	contains 0.49 zinc atoms per subunit	Mycobacterium tuberculosis
Zn2+	contains 0.83 zinc atoms per subunit	Pyricularia grisea
Zn2+	contains 0.97 zinc atoms per subunit	Bacillus cereus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
30540	-	1 * 30671 + 1 * 30540, mass spectrometry	Bacillus cereus
30671	-	1 * 30671 + 1 * 30540, mass spectrometry	Bacillus cereus
36411	-	4 * 36411, mass spectrometry	Mycobacterium tuberculosis
38441	-	4 * 38441, mass spectrometry	Pseudomonas aeruginosa
39644	-	1 * 39775 + 1 * 39644, mass spectrometry	Pyricularia grisea
39775	-	1 * 39775 + 1 * 39644, mass spectrometry	Pyricularia grisea

Organic Solvent Stability

Organic Solvent	Comment	Organism
Acetone	more than 50% of the enzyme activity is retained in the presence of 10% (v/v) acetone	Pyricularia grisea
Acetone	more than 50% of the enzyme activity is retained in the presence of 25% (v/v) acetone	Pseudomonas aeruginosa
Acetone	more than 50% of the enzyme activity is retained in the presence of 25% (v/v) acetone	Bacillus cereus
Acetone	more than 50% of the enzyme activity is retained in the presence of 25% (v/v) acetone	Mycobacterium tuberculosis
acetonitrile	more than 50% of the enzyme activity is retained in the presence of 10% (v/v) acetonitrile	Bacillus cereus
acetonitrile	more than 50% of the enzyme activity is retained in the presence of 10% (v/v) acetonitrile	Mycobacterium tuberculosis
acetonitrile	more than 50% of the enzyme activity is retained in the presence of 15% (v/v) acetonitrile	Pseudomonas aeruginosa
acetonitrile	more than 50% of the enzyme activity is retained in the presence of about 5% (v/v) acetonitrile	Pyricularia grisea
dimethylformamide	more than 50% of the enzyme activity is retained in the presence of 10% (v/v) dimethylformamide	Pyricularia grisea
dimethylformamide	more than 50% of the enzyme activity is retained in the presence of 20% (v/v) dimethylformamide	Mycobacterium tuberculosis
dimethylformamide	more than 50% of the enzyme activity is retained in the presence of 35% (v/v) dimethylformamide	Pseudomonas aeruginosa
dimethylformamide	more than 50% of the enzyme activity is retained in the presence of 50% (v/v) dimethylformamide	Bacillus cereus
DMSO	167% activity in the presence of 50% (v/v) DMSO	Bacillus cereus
DMSO	more than 50% of the enzyme activity is retained in the presence of 20% (v/v) DMSO	Pyricularia grisea
DMSO	more than 50% of the enzyme activity is retained in the presence of 50% (v/v) DMSO	Pseudomonas aeruginosa
DMSO	more than 50% of the enzyme activity is retained in the presence of 50% (v/v) DMSO	Mycobacterium tuberculosis
tert-Butanol	more than 50% of the enzyme activity is retained in the presence of 15% (v/v) tert-butanol	Pseudomonas aeruginosa
tert-Butanol	more than 50% of the enzyme activity is retained in the presence of 20% (v/v) tert-butanol	Mycobacterium tuberculosis
tert-Butanol	more than 50% of the enzyme activity is retained in the presence of about 5% (v/v) tert-butanol	Bacillus cereus
tert-Butanol	more than 50% of the enzyme activity is retained in the presence of about 5% (v/v) tert-butanol	Pyricularia grisea

Organism

Organism	UniProt	Comment	Textmining
Bacillus cereus	-	-	-
Bacillus cereus ATCC 10987	-	-	-
Mycobacterium tuberculosis	-	-	-
Pseudomonas aeruginosa	-	-	-
Pyricularia grisea	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
ammonium sulfate precipitation, DEAE Sepharose column chromatography, and Sepharose Q column chromatography	Pseudomonas aeruginosa
ammonium sulfate precipitation, DEAE Sepharose column chromatography, and Sepharose Q column chromatography	Bacillus cereus
ammonium sulfate precipitation, DEAE Sepharose column chromatography, and Sepharose Q column chromatography	Mycobacterium tuberculosis
ammonium sulfate precipitation, DEAE Sepharose column chromatography, and Sepharose Q column chromatography	Pyricularia grisea

Storage Stability

Storage Stability	Organism
4°C, in 50 mM HEPES buffer with pH 7.3 supplemented with ZnCl2, 24 h, 40% loss of activity	Bacillus cereus
4°C, in 50 mM HEPES buffer with pH 7.3 supplemented with ZnCl2, several days, the enzyme remains stable	Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
D-fructose 1,6-bisphosphate	-	Pseudomonas aeruginosa	dihydroxyacetone phosphate + D-glyceraldehyde 3-phosphate	-	r
D-fructose 1,6-bisphosphate	-	Bacillus cereus	dihydroxyacetone phosphate + D-glyceraldehyde 3-phosphate	-	r
D-fructose 1,6-bisphosphate	-	Mycobacterium tuberculosis	dihydroxyacetone phosphate + D-glyceraldehyde 3-phosphate	-	r
D-fructose 1,6-bisphosphate	-	Pyricularia grisea	dihydroxyacetone phosphate + D-glyceraldehyde 3-phosphate	-	r
D-fructose 1,6-bisphosphate	-	Bacillus cereus ATCC 10987	dihydroxyacetone phosphate + D-glyceraldehyde 3-phosphate	-	r

Subunits

Subunits	Comment	Organism
heterodimer	1 * 30671 + 1 * 30540, mass spectrometry	Bacillus cereus
heterodimer	1 * 39775 + 1 * 39644, mass spectrometry	Pyricularia grisea
homotetramer	4 * 36411, mass spectrometry	Mycobacterium tuberculosis
homotetramer	4 * 38441, mass spectrometry	Pseudomonas aeruginosa

Synonyms

Synonyms	Comment	Organism
class II fructose 1,6-bisphosphate aldolase	-	Pseudomonas aeruginosa
class II fructose 1,6-bisphosphate aldolase	-	Bacillus cereus
class II fructose 1,6-bisphosphate aldolase	-	Mycobacterium tuberculosis
class II fructose 1,6-bisphosphate aldolase	-	Pyricularia grisea
FBP aldolase	-	Pseudomonas aeruginosa
FBP aldolase	-	Bacillus cereus
FBP aldolase	-	Mycobacterium tuberculosis
FBP aldolase	-	Pyricularia grisea

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
45	60	the aldolase loses 25% of its activity after 10 min of incubation at 45°C and loses almost all its activity after a10 min incubation at 60°C	Pyricularia grisea
60	-	the aldolase loses almost all its activity after a 10 min incubation at 60°C	Mycobacterium tuberculosis
60	-	the aldolase loses almost all its activity after a10 min incubation at 60°C	Pseudomonas aeruginosa
90	-	the aldolase retains almost half of its activity after 10 min of incubation at 90°C	Bacillus cereus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
1.53	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Pseudomonas aeruginosa
2.95	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Bacillus cereus
30.1	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Mycobacterium tuberculosis
45.7	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Pyricularia grisea

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.8	-	-	Mycobacterium tuberculosis
7.8	-	-	Pyricularia grisea
8	-	-	Bacillus cereus
8.5	-	-	Pseudomonas aeruginosa

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.066	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Bacillus cereus
0.44	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Pseudomonas aeruginosa
9	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Pyricularia grisea
10.8	-	D-fructose 1,6-bisphosphate	in Tris-HCl pH 8.0, at 30°C	Mycobacterium tuberculosis