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Literature summary extracted from

  • Angelov, A.; Putyrski, M.; Liebl, W.
    Molecular and biochemical characterization of alpha-glucosidase and alpha-mannosidase and their clustered genes from the thermoacidophilic archaeon Picrophilus torridus (2006), J. Bacteriol., 188, 7123-7131.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.20 gene aglA, expression analysis and mapping of transcription start sites, the gene is clustered in the genome and shows colinear orientation and a small intergenic space with gene manA encoding alpha-mannosidase, the genes are independently transcribed, both producing leaderless mRNA, phylogenetic tree, overexpression in Escherichia coli strain BL21, subcloning in Escherichia coli strain XL-1 Blue Picrophilus torridus
3.2.1.24 overexpressed in Escherichia coli Picrophilus torridus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.24 Ag+ strong inhibitor at concentrations of 1 mM or 10 mM Picrophilus torridus
3.2.1.24 Cu2+ strong inhibitor at concentrations of 1 mM or 10 mM Picrophilus torridus
3.2.1.24 EDTA supplied in equimolar amounts or in excess, neutralizes also effect of resistance of ManA to heat inactivation Picrophilus torridus
3.2.1.24 Fe3+ strong inhibitor at concentrations of 1 mM or 10 mM Picrophilus torridus
3.2.1.24 additional information 1-deoxymannojirimycin has no effect on ManA at concentrations up to 500 microM Picrophilus torridus
3.2.1.24 Ni2+ strong inhibitor at concentrations of 1 mM or 10 mM Picrophilus torridus
3.2.1.24 swainsonine shows competitive inhibition when p-nitrophenyl-Man is used as a substrate Picrophilus torridus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.24 Cd2+ potent activator at concentration of 1 mM, 12.5fold activation compared to the apoenzyme Picrophilus torridus
3.2.1.24 Co2+ 8.3fold activation at a concentration of 10 mM, resistance of ManA to heat inactivation is increased by presence of Co2+ Picrophilus torridus
3.2.1.24 Fe2+ 4.5fold activation at a concentration of 10 mM Picrophilus torridus
3.2.1.24 Mn2+ 3.8fold activation at a concentration of 10 mM Picrophilus torridus
3.2.1.24 additional information Zn2+, Ca2+, Mg2+, Ba2+, K+, and Li+ had no significant effect on ManA activity at a concentration of 1 mM or 10 mM Picrophilus torridus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.20 72000
-
 6 * 72000, recombinant AglA, SDS-PAGE Picrophilus torridus
3.2.1.20 440000
-
 recombinant AglA, gel filtration Picrophilus torridus
3.2.1.24 111500
-
 2 * 111500, gel filtration Picrophilus torridus
3.2.1.24 120000
-
 small fraction of ManA activity, reflecting single subunits Picrophilus torridus
3.2.1.24 220000
-
 gel filtration Picrophilus torridus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.20 maltose + H2O Picrophilus torridus probable physiological role of the enzyme in the utilization of exogenous glycoproteins and/or in the turnover of the organism’s own glycoproteins, overview alpha-D-glucose + D-glucose
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.20 Picrophilus torridus Q6L2X4 strain DSM 9790, gene aglA
-
3.2.1.24 Picrophilus torridus Q6L2X5 extremophilic organism belonging to the euryarchaeal phylum
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.20 recombinant AglA 39.3fold from Escherichia coli strain BL21 by heat treatment for 15 min at 67°C, hydrophobic interaction and anion exchange chromatography, and gel filtration in presence of 150 mM NaCl Picrophilus torridus
3.2.1.24 SDS-PAGE, anion-exchange, size exclusion chromatography Picrophilus torridus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.1.20 additional information 60°C is the optimal, 67°C the maximal growth temperature Picrophilus torridus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.24 6.6
-
 ManA is equally active with mannobiose substrates, released mannose is quantified, resulting in a specific activity of approximately 6.6 U/mg Picrophilus torridus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.20 4-nitrophenyl alpha-D-glucopyranoside + H2O low activity Picrophilus torridus 4-nitrophenol + alpha-D-glucose
-
 ?
3.2.1.20 dextrin 10 + H2O 27% of the activity with maltose Picrophilus torridus alpha-D-glucose
-
 ?
3.2.1.20 maltohexaose + H2O
-
 Picrophilus torridus maltopentaose + D-glucose
-
 ?
3.2.1.20 maltopentaose + H2O
-
 Picrophilus torridus alpha-D-glucose + ?
-
 ?
3.2.1.20 maltose + H2O probable physiological role of the enzyme in the utilization of exogenous glycoproteins and/or in the turnover of the organism’s own glycoproteins, overview Picrophilus torridus alpha-D-glucose + D-glucose
-
 ?
3.2.1.20 maltose + H2O preferred substrate, the enzyme shows strict glucose substrate specificity hydrolyzing maltose, as well as longer alpha-1,4-linked maltooligosaccharides Picrophilus torridus alpha-D-glucose + D-glucose
-
 ?
3.2.1.20 maltotetraose + H2O 54% of the activity with maltose Picrophilus torridus alpha-D-glucose
-
 ?
3.2.1.20 maltotriose + H2O
-
 Picrophilus torridus maltose + alpha-D-glucose
-
 ?
3.2.1.24 1,2-alpha-D-mannobiose + H2O
-
 Picrophilus torridus alpha-D-mannose
-
 ?
3.2.1.24 1,3-alpha-D-mannobiose + H2O
-
 Picrophilus torridus alpha-D-mannose
-
 ?
3.2.1.24 1,6-alpha-D-mannobiose + H2O
-
 Picrophilus torridus alpha-D-mannose
-
 ?
3.2.1.24 alpha3,alpha6-D-mannopentaose + H2O
-
 Picrophilus torridus alpha-D-mannose
-
 ?
3.2.1.24 methyl-O-1,4-alpha-D-mannobiose + H2O
-
 Picrophilus torridus alpha-D-mannose
-
 ?
3.2.1.24 p-nitrophenyl-alpha-D-mannopyranoside + H2O
-
 Picrophilus torridus 4-nitrophenol + alpha-D-mannose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.20 hexamer 6 * 72000, recombinant AglA, SDS-PAGE Picrophilus torridus
3.2.1.24 dimer 2 * 111500, gel filtration Picrophilus torridus

Synonyms

EC Number Synonyms Comment Organism
3.2.1.20 AglA
-
 Picrophilus torridus
3.2.1.24 alpha-mannosidase
-
 Picrophilus torridus
3.2.1.24 ManA
-
 Picrophilus torridus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.20 87
-
 recombinant AglA Picrophilus torridus
3.2.1.24 70
-
 maximum activity of ManA Picrophilus torridus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.20 100
-
 15 min, inactivation Picrophilus torridus
3.2.1.24 80
-
 resistance of ManA to heat inactivation (at 80°C) Picrophilus torridus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.20 5
-
-
 Picrophilus torridus
3.2.1.24 5.2
-
 maximum activity of ManA Picrophilus torridus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.24 0.00122
-
 swainsonine
-
 Picrophilus torridus