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Literature summary for 3.2.1.B36 extracted from

  • Park, K.H.; Jung, J.H.; Park, S.G.; Lee, M.E.; Holden, J.F.; Park, C.S.; Woo, E.J.
    Structural features underlying the selective cleavage of a novel exo-type maltose-forming amylase from Pyrococcus sp. ST04 (2014), Acta Crystallogr. Sect. D, 70, 1659-1668.
    View publication on PubMed
Search for more literature for 3.2.1.B36

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of selenomethionine-labeled wild-type and mutant enzymes in Escherichia coli strain BL21-CodonPlus(DE3)-RP Pyrococcus sp.

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant selenomethionine-labeled wild-type enzyme, sitting drop vapour diffusion method, mixing of 0.001 ml of 15 mg/ml protein solution with 0.001 ml of reservoir solution consisting of 1.0 M ammonium citrate dibasic, 0.8 M sodium acetate trihydrate pH 4.6, 18°C, X-ray diffraction structure determination and analysis at 1..8 A resolution Pyrococcus sp.

Protein Variants

Protein Variants Comment Organism
D253N site-directed mutagenesis, the mutant shows reduced alpha-1,4- and alpha1,6-hydrolytic activity compared to the wild-type enzyme Pyrococcus sp.
E153Q site-directed mutagenesis, the mutant shows reduced alpha-1,4- and alpha1,6-hydrolytic activity compared to the wild-type enzyme Pyrococcus sp.
E580Q site-directed mutagenesis, the mutant shows reduced alpha-1,4- and alpha1,6-hydrolytic activity compared to the wild-type enzyme Pyrococcus sp.
F218A site-directed mutagenesis, the mutant exhibits 3.5fold icreased alpha-1,4-glucosidic bond hydrolysis compared to the wild-type enzyme Pyrococcus sp.
F452A site-directed mutagenesis, the mutant shows reduced alpha-1,4- and alpha1,6-hydrolytic activity compared to the wild-type enzyme Pyrococcus sp.
W453A site-directed mutagenesis, the mutant shows reduced alpha-1,4- and alpha1,6-hydrolytic activity compared to the wild-type enzyme Pyrococcus sp.

Metals/Ions

Metals/Ions Comment Organism Structure
additional information no metal ions Pyrococcus sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
70191
-
 2 * 70191, recombinant enzyme, SDS-PAGE, in the dimeric structure, extensive interactions including Asn185-Gly327, Glu224-Lys241 and Gly327-Asn185 hydrogen bonds and a Glu224-Lys241 salt bridge between two (beta/alpha)7-barrels of the N-domains comprise the dimer associations Pyrococcus sp.
260000
-
 recombinant enzyme, gel filtration Pyrococcus sp.

Organism

Organism UniProt Comment Textmining
Pyrococcus sp. I3RE04
-
-
Pyrococcus sp. ST04 I3RE04
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21-CodonPlus(DE3)-RP Pyrococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the enzyme hydrolyzes both alpha-1,4-glucosidic and alpha-1,6-glucosidic linkages of substrates, recognizing only maltose units, in an exo-type manner. Substrate specificity and binding, docking modelling, overview Pyrococcus sp. ?
-
 ?
additional information the enzyme hydrolyzes both alpha-1,4-glucosidic and alpha-1,6-glucosidic linkages of substrates, recognizing only maltose units, in an exo-type manner. Substrate specificity and binding, docking modelling, overview Pyrococcus sp. ST04 ?
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 70191, recombinant enzyme, SDS-PAGE, in the dimeric structure, extensive interactions including Asn185-Gly327, Glu224-Lys241 and Gly327-Asn185 hydrogen bonds and a Glu224-Lys241 salt bridge between two (beta/alpha)7-barrels of the N-domains comprise the dimer associations Pyrococcus sp.
More the enzyme structure shows a tight ring-shaped tetramer with monomers composed of two domains: an N-domain (amino acids 1-341) with a typical GH57 family (beta/alpha)7-barrel fold and a C-domain (amino acids 342-597) composed of alpha-helical bundles, two domains assemble to form a groove in the active site Pyrococcus sp.

Synonyms

Synonyms Comment Organism
maltose-forming alpha-amylase
-
 Pyrococcus sp.
PSMA
-
 Pyrococcus sp.

General Information

General Information Comment Organism
evolution the enzyme belongs to the glycoside hydrolase family 57, GH57 Pyrococcus sp.
additional information the enzyme structure shows a tight ring-shaped tetramer with monomers composed of two domains: an N-domain (amino acids 1-341) with a typical GH57 family (beta/alpha)7-barrel fold and a C-domain (amino acids 342-597) composed of alpha-helical bundles. The catalytic residues are Glu153 and Asp253 at the domain interface Pyrococcus sp.