Literature summary for 3.1.3.8 extracted from

Tan, H.; Wu, X.; Xie, L.; Huang, Z.; Peng, W.; Gan, B.
Identification and characterization of a mesophilic phytase highly resilient to high-temperatures from a fungus-garden associated metagenome (2016), Appl. Microbiol. Biotechnol., 100, 2225-2241 .

Activating Compound

Activating Compound	Comment	Organism
CTAB	20% activation at 1% w/v	Dendroctonus frontalis
glycerol	activates by 38% at 10% v/v	Dendroctonus frontalis
additional information	no effect by EDTA at 0.1-20 mM	Dendroctonus frontalis
Tween 20	about 22% and 30% activation at 0.5% and 1% v/v, respectively	Dendroctonus frontalis
Tween 80	about 27% and 35% activation at 0.5% and 1% v/v, respectively	Dendroctonus frontalis

Cloned(Commentary)

Cloned (Comment)	Organism
gene phyTX52, phylogenetic analysis, recombinant overexpression of wild-type enzyme in Escherichia coli strain BL21(DE3)	Dendroctonus frontalis

Protein Variants

Protein Variants	Comment	Organism
E378A/D128A/D71A/E443A	site-directed mutagenesis, mutant rPhyXT52:DELTASB1,3,6,7 with loss of four salt bridges	Dendroctonus frontalis
E378A/D178A	site-directed mutagenesis, mutant rPhyXT52:DELTASB1,5 with loss of two salt bridges	Dendroctonus frontalis
E378A/R56A/D128A/D267A/D178A/D71A/E443A	site-directed mutagenesis, mutant rPhyXT52:DELTASB1-7 with loss of seven salt bridges	Dendroctonus frontalis
E378A/R56A/D267A/E443A	site-directed mutagenesis, mutant rPhyXT52:DELTASB1,2,4,7 with loss of four salt bridges	Dendroctonus frontalis
additional information	construction of mutants that lose salt bridges through the mutations, the combinations of the disabled salt bridges are randomly selected. The mutants show reduced melting temperatures and reduced optimum reaction temperature compared to wild-type rPhyXT52, overview	Dendroctonus frontalis
R56A/D128A	site-directed mutagenesis, mutant rPhyXT52:DELTASB2,3 with loss of two salt bridges	Dendroctonus frontalis

Inhibitors

Inhibitors	Comment	Organism
2-mercaptoethanol	20% inhibition at 1 mM, 66% at 5 mM	Dendroctonus frontalis
Ag+	about 40% inhibition at 1 mM, 82% at 5 mM	Dendroctonus frontalis
Al3+	67% inhibition at 1 mM, 88% at 5 mM	Dendroctonus frontalis
Ba2+	about 60% inhibition at 1 mM, 83% at 5 mM	Dendroctonus frontalis
Ca2+	slightly elevates the phytase activity at 20 mM but inhibits the activity at a higher concentration of 50 mM	Dendroctonus frontalis
Cd2+	92.5% inhibition at 1 mM	Dendroctonus frontalis
citrate	slight inhibition at 20-100 mM	Dendroctonus frontalis
Co2+	about 69% inhibition at 1 mM, 83% at 5 mM	Dendroctonus frontalis
Cr3+	87% inhibition at 1 mM	Dendroctonus frontalis
Cu2+	72% inhibition at 5 mM	Dendroctonus frontalis
DTT	23% inhibition at 1 mM, 88% at 5 mM	Dendroctonus frontalis
Fe2+	9% inhibition at 5 mM	Dendroctonus frontalis
glycerol	inhibits by 14% at 20% v/v	Dendroctonus frontalis
Mg2+	has no significant influence at low concentrations of 1 and 5 mM and inhibits the phytase activity at high concentrations of 20 and 50 mM	Dendroctonus frontalis
Mn2+	67% inhibition at 5 mM	Dendroctonus frontalis
additional information	no effect by EDTA at 0.1-20 mM	Dendroctonus frontalis
Ni2+	about 55% inhibition at 1 mM, 78% at 5 mM	Dendroctonus frontalis
Pb2+	94% inhibition at 1 mM, almost complete inhibition at 5 mM	Dendroctonus frontalis
PMSF	44% inhibition at 2 mM	Dendroctonus frontalis
SDS	73% inhibition at 0.1% w/v, 85% at 0.5% w/v, and 94% at 2% w/v	Dendroctonus frontalis
Triton-X-100	12% inhibition at 0.5% v/v, 37% at 1% v/v	Dendroctonus frontalis
Urea	30% inhibition at 2 M, 44% at 4 M	Dendroctonus frontalis
Zn2+	53% inhibition at 5 mM	Dendroctonus frontalis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	Michaelis-Menten kinetics	Dendroctonus frontalis
0.262	-	myo-inositol hexakisphosphate	recombinant enzyme, pH 3.9, 52.5°C	Dendroctonus frontalis
0.326	-	myo-inositol hexakisphosphate	recombinant enzyme, pH 3.9, 37°C	Dendroctonus frontalis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Ca2+	slightly elevates the phytase activity at 20 mM but inhibits the activity at a higher concentration of 50 mM	Dendroctonus frontalis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
51700	-	recombinant enzyme, gel filtration	Dendroctonus frontalis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
myo-inositol hexakisphosphate + H2O	Dendroctonus frontalis	highly preferred substrate	1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Dendroctonus frontalis	KM873028	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli strain BL21(DE3)	Dendroctonus frontalis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
4175	-	purified recombinant enzyme, pH 3.9, 52.5°C	Dendroctonus frontalis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	enzyme rPhysXT52 shows nonspecific phosphohydrolytic activity with 4-nitrophenyl phosphate, 1-naphthyl phosphate, 2-naphthyl phosphate, phenyl phosphate, 2-glycerophosphate, glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-diphosphate, AMP, ADP, ATP, and NADP+, overview	Dendroctonus frontalis	?	-	?
myo-inositol hexakisphosphate + H2O	-	Dendroctonus frontalis	1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate	-	?
myo-inositol hexakisphosphate + H2O	highly preferred substrate	Dendroctonus frontalis	1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate	-	?

Subunits

Subunits	Comment	Organism
monomer	1 * 52000, recombinant enzyme, SDS-PAGE	Dendroctonus frontalis

Synonyms

Synonyms	Comment	Organism
PhyTX52	-	Dendroctonus frontalis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
52.5	-	-	Dendroctonus frontalis

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
60	100	purified recombinant enzyme, half-life at 60, 80, and 100°C is approximate 5.6 h, 2.1 h, and 27 min, respectively. At 80°C, recombinant PhyXT52 retains about 90% residual activity after 20 min, and at 100°C more than 93% residual activity are retained after 15 min and over 83% after 20 min. Melting temperatures of wild-type rPhyXT52 and mutant enzymes rPhyXT52:DELTASB1,5, rPhyXT52:DELTASB2,3, rPhyXT52:DELTASB1,2,4,7, rPhyXT52:DELTASB1,3,6,7, rPhyXT52:DELTASB1-7 are 60.5°C, 58.2°C; 59.7°C; 54.6°C, 55.2°C, and 52.8°C. Half-lifes of mutants are determined at 60-100°C, overview	Dendroctonus frontalis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
284600	-	myo-inositol hexakisphosphate	recombinant enzyme, pH 3.9, 37°C	Dendroctonus frontalis
375200	-	myo-inositol hexakisphosphate	recombinant enzyme, pH 3.9, 52.5°C	Dendroctonus frontalis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
3.9	-	recombinant enzyme	Dendroctonus frontalis

pH Range

pH Minimum	pH Maximum	Comment	Organism
3	6	activity range, mesophilic phytase, profile overview	Dendroctonus frontalis

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the histidine acid phosphatase (HAP) family phytases present in insect-cultivated fungus gardens	Dendroctonus frontalis
additional information	structural model of rPhyXT52 is deduced by homology modeling using the Yersinia kristensenii phytase (PDB ID 4ARV) structure as template, overview	Dendroctonus frontalis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
873006	-	myo-inositol hexakisphosphate	recombinant enzyme, pH 3.9, 37°C	Dendroctonus frontalis
1432060	-	myo-inositol hexakisphosphate	recombinant enzyme, pH 3.9, 52.5°C	Dendroctonus frontalis