Literature summary for 3.6.1.11 extracted from

Zhang, A.; Guo, E.; Qian, L.; Tang, N.Y.; Watt, R.M.; Bartlam, M.
Purification, crystallization and X-ray crystallographic analysis of a putative exopolyphosphatase from Zymomonas mobilis (2016), Acta Crystallogr. Sect. F, 72, 172-178 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Zymomonas mobilis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant detagged wild-type enzyme by hanging drop vapour diffusion, mixing of 0.002 ml of 10 mg/ml protein in 50 mM Tris, pH 8.0, 500 mM NaCl, and 5% glycerol, with 0.001 ml of reservoir solution containing 15.2% w/v PEG 3350, 2% w/v PEG 8000, 72 mM Tris, pH 8.0, 144 mM magnesium chloride hexahydrate, 20 mM HEPES/sodium hydroxide pH 7.5, 1.6% v/v glycerol, and 8% v/v DMSO, and equilibration against 0.2 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 3.3 A resolution. Purified recombinant detagged mutant truncated enzyme and truncated mutant E137A by sitting drop vapour diffusion, mixing of 0.001 ml of 8 mg/ml protein in 50 mM Tris, pH 8.0, 500 mM NaCl, and 5% glycerol, with 0.001 ml of reservoir solution containing 0.2 M magnesium chloride hexahydrate, 0.1 M Tris, pH 8.5 or pH 6.5, respectively, 25% w/v PEG 3350, and equilibration against 0.1 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 1.80 and 1.56 A resolution, respectively. Molecular replacement	Zymomonas mobilis

Crystallization (Comment)

Organism

purified recombinant detagged wild-type enzyme by hanging drop vapour diffusion, mixing of 0.002 ml of 10 mg/ml protein in 50 mM Tris, pH 8.0, 500 mM NaCl, and 5% glycerol, with 0.001 ml of reservoir solution containing 15.2% w/v PEG 3350, 2% w/v PEG 8000, 72 mM Tris, pH 8.0, 144 mM magnesium chloride hexahydrate, 20 mM HEPES/sodium hydroxide pH 7.5, 1.6% v/v glycerol, and 8% v/v DMSO, and equilibration against 0.2 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 3.3 A resolution. Purified recombinant detagged mutant truncated enzyme and truncated mutant E137A by sitting drop vapour diffusion, mixing of 0.001 ml of 8 mg/ml protein in 50 mM Tris, pH 8.0, 500 mM NaCl, and 5% glycerol, with 0.001 ml of reservoir solution containing 0.2 M magnesium chloride hexahydrate, 0.1 M Tris, pH 8.5 or pH 6.5, respectively, 25% w/v PEG 3350, and equilibration against 0.1 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 1.80 and 1.56 A resolution, respectively. Molecular replacement

Zymomonas mobilis

Protein Variants

Protein Variants	Comment	Organism
E137A	site-directed mutagenesis of the truncated mutant ZmPPX(30-508), an active site mutant	Zymomonas mobilis
additional information	truncation of 29 amino acids from the N-terminus of the enzyme protein of mutant ZmPPX(30-508) results in a significant improvement in the diffraction of ZmPPX crystals from 3.3 to 1.8 A resolution using synchrotron radiation	Zymomonas mobilis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Zymomonas mobilis

Organism

Organism	UniProt	Comment	Textmining
Zymomonas mobilis	A0A542W0V0	-	-
Zymomonas mobilis NCIMB 11163	A0A542W0V0	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration, followed by cleavage of the His tag by PreScission protease, and gel filtration	Zymomonas mobilis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	recombinant ZmPPX possesses exopolyphosphatase activity against a synthetic poly-P substrate	Zymomonas mobilis	?	-	-
additional information	recombinant ZmPPX possesses exopolyphosphatase activity against a synthetic poly-P substrate	Zymomonas mobilis NCIMB 11163	?	-	-
polyphosphate130 + H2O	-	Zymomonas mobilis	polyphosphate129 + phosphate	-	?
polyphosphate130 + H2O	-	Zymomonas mobilis NCIMB 11163	polyphosphate129 + phosphate	-	?

Subunits

Subunits	Comment	Organism
dimer	analytical ultracentrifugation	Zymomonas mobilis

Synonyms

Synonyms	Comment	Organism
PPX	-	Zymomonas mobilis
Za10_0559	locus name	Zymomonas mobilis
ZmPPX	-	Zymomonas mobilis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Zymomonas mobilis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.8	-	assay at	Zymomonas mobilis

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the PPX/GppA phosphatase family (pfam02541) that consists of PPX (EC 3.6.1.11) and guanosine pentaphosphate phosphohydrolase (GppA, EC 3.6.1.40) enzymes	Zymomonas mobilis
metabolism	polyphosphate kinase (PPK) is the principal source of polyphosphate in most bacteria, whereas exopolyphosphatases (PPX) are mainly responsible for the degradation of polyphosphate	Zymomonas mobilis
physiological function	exopolyphosphatase (PPX) enzymes degrade inorganic polyphosphate (poly-P), which is essential for the survival of microbial cells in response to external stresses. Inorganic polyphosphate (poly-P), comprising a few to hundreds of orthophosphate residues linked by high-energy phosphoanhydride bonds, is found in virtually all living cells	Zymomonas mobilis