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Literature summary for 3.2.1.1 extracted from

  • Jung, J.H.; An, Y.K.; Son, S.Y.; Jeong, S.Y.; Seo, D.H.; Kim, M.K.; Park, C.S.
    Characterization of a novel extracellular alpha-amylase from Ruminococcus bromii ATCC 27255 with neopullulanase-like activity (2019), Int. J. Biol. Macromol., 130, 605-614 .
    View publication on PubMed
Search for more literature for 3.2.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21 (DE3) Ruminococcus bromii
gene RB5AMG_01539, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21-CodonPlus (DE3)-RP Ruminococcus bromii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetics with different substrates, overview Ruminococcus bromii
additional information
-
 pullulan Km-value: 15.4 mg/ml, 45°C, pH 5.0 Ruminococcus bromii
additional information
-
 starch Km-value: 16.0 mg/ml, 45°C, pH 5.0 Ruminococcus bromii
additional information
-
 amylose Km-value: 6.17 mg/ml, 45°C, pH 5.0 Ruminococcus bromii
additional information
-
 amylopectin Km-value: 8.95 mg/ml, 45°C, pH 5.0 Ruminococcus bromii

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
 Ruminococcus bromii
-
-

Organism

Organism UniProt Comment Textmining
Ruminococcus bromii
-
-
-
Ruminococcus bromii A0A2N0UQI0
-
-
Ruminococcus bromii ATCC 27255
-
-
-
Ruminococcus bromii ATCC 27255 A0A2N0UQI0
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Ruminococcus bromii
recombinant His6-tagged enzyme from Escherichia coli strain BL21-CodonPlus (DE3)-RP by nickel affinity chromatography and ultrafiltration Ruminococcus bromii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
46.1
-
 purified recombinant His-tagged enzyme, pH 5.0, 45°C, substrate amylopectin Ruminococcus bromii
72.5
-
 45°C, pH 5.0 Ruminococcus bromii
72.5
-
 purified recombinant His-tagged enzyme, pH 5.0, 45°C, substrate pullulan Ruminococcus bromii
254
-
 purified recombinant His-tagged enzyme, pH 5.0, 45°C, substrate soluble starch Ruminococcus bromii
384
-
 purified recombinant His-tagged enzyme, pH 5.0, 45°C, substrate amylose Ruminococcus bromii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
amylopectin + H2O at 12% of the activity as compared to amylose Ruminococcus bromii ?
-
 ?
amylopectin + H2O at 12% of the activity as compared to amylose Ruminococcus bromii ATCC 27255 ?
-
 ?
amylopectin + H2O 0.5% substrate solution, 12% activity compared to amylose Ruminococcus bromii maltose + D-glucose
-
 ?
amylose + H2O
-
 Ruminococcus bromii ?
-
 ?
amylose + H2O
-
 Ruminococcus bromii ATCC 27255 ?
-
 ?
amylose + H2O 0.5% substrate solution, best substrate Ruminococcus bromii maltose + D-glucose
-
 ?
amylose + H2O 0.5% substrate solution, best substrate Ruminococcus bromii ATCC 27255 maltose + D-glucose
-
 ?
gamma-cyclodextrin + H2O at 3.9% of the activity as compared to amylose Ruminococcus bromii ?
-
 ?
gamma-cyclodextrin + H2O at 3.9% of the activity as compared to amylose Ruminococcus bromii ATCC 27255 ?
-
 ?
maltopentaose + H2O
-
 Ruminococcus bromii maltose + ?
-
 ?
maltopentaose + H2O
-
 Ruminococcus bromii maltotriose + maltose + D-glucose
-
 ?
maltotetraose + H2O
-
 Ruminococcus bromii 2 maltose
-
 ?
maltotetraose + H2O
-
 Ruminococcus bromii ATCC 27255 2 maltose
-
 ?
maltotetraose + H2O
-
 Ruminococcus bromii maltose + ?
-
 ?
maltotriose + H2O
-
 Ruminococcus bromii maltose + D-glucose
-
 ?
maltotriose + H2O
-
 Ruminococcus bromii ATCC 27255 maltose + D-glucose
-
 ?
maltotriose + H2O
-
 Ruminococcus bromii maltose + ?
-
 ?
additional information alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, and malto-beta-cyclodextrin are poor substrates. Thin layer chromatography and high performance anion exchange chromatography analysis of hydrolysis products from maltodextrins (G2-G5) by recombinant enzyme Rbamy5, overview. Maltose is not hydrolyzed Ruminococcus bromii ?
-
 ?
additional information alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, and malto-beta-cyclodextrin are poor substrates. Thin layer chromatography and high performance anion exchange chromatography analysis of hydrolysis products from maltodextrins (G2-G5) by recombinant enzyme Rbamy5, overview. Maltose is not hydrolyzed Ruminococcus bromii ATCC 27255 ?
-
 ?
pullulan + H2O 0.5% substrate solution, 18.9% activity compared to amylose Ruminococcus bromii panose + ?
-
 ?
pullulan + H2O at 72.5% of the activity as compared to amylose. The enzyme is an alpha-amylase with neopullulanase-like activity Ruminococcus bromii panose + ?
-
 ?
pullulan + H2O at 72.5% of the activity as compared to amylose. The enzyme is an alpha-amylase with neopullulanase-like activity Ruminococcus bromii ATCC 27255 panose + ?
-
 ?
pullulan + H2O 0.5% substrate solution, 18.9% activity compared to amylose Ruminococcus bromii ATCC 27255 panose + ?
-
 ?
soluble starch + H2O 0.5% substrate solution, 66.2% activity compared to amylose Ruminococcus bromii maltose + D-glucose
-
 ?
starch + H2O at 66.2% of the activity as compared to amylose Ruminococcus bromii ?
-
 ?
starch + H2O at 66.2% of the activity as compared to amylose Ruminococcus bromii ATCC 27255 ?
-
 ?

Subunits

Subunits Comment Organism
? x * 60000, recombinant His6-tagged enzyme, SDS-PAGE, x * 58500, about, sequence calculation Ruminococcus bromii

Synonyms

Synonyms Comment Organism
amy5
-
 Ruminococcus bromii
RB5AMG_01539
-
 Ruminococcus bromii
Rbamy5
-
 Ruminococcus bromii
Ruminococcus bromii intermediary alpha-amylase 5
-
 Ruminococcus bromii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 recombinant enzyme Ruminococcus bromii
45
-
 substrate: soluble starch Ruminococcus bromii

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 50 30°C: about 40% of maximal activity, 50°C: about 40% of maximal activity, substrate: soluble starch Ruminococcus bromii
30 50 40% of maximal activity at 30°C and 50°C, profile overview Ruminococcus bromii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
35
-
 140 min, no loss of activity Ruminococcus bromii
35
-
 purified recombinant enzyme, completely stable for 140 min Ruminococcus bromii
40
-
 120 min, about 45% loss of activity Ruminococcus bromii
40
-
 purified recombinant enzyme, loss of 50% activity after 140 min Ruminococcus bromii
45
-
 20 min, about 70% loss of activity Ruminococcus bromii
45
-
 purified recombinant enzyme, inactivation within about 27 min Ruminococcus bromii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
137
-
 amylopectin 45°C, pH 5.0 Ruminococcus bromii
137
-
 amylopectin recombinant His-tagged enzyme, pH 5.0, 45°C Ruminococcus bromii
297
-
 pullulan 45°C, pH 5.0 Ruminococcus bromii
297
-
 pullulan recombinant His-tagged enzyme, pH 5.0, 45°C Ruminococcus bromii
1014
-
 amylose 45°C, pH 5.0 Ruminococcus bromii
1014
-
 amylose recombinant His-tagged enzyme, pH 5.0, 45°C Ruminococcus bromii
1055
-
 starch 45°C, pH 5.0 Ruminococcus bromii
1055
-
 soluble starch recombinant His-tagged enzyme, pH 5.0, 45°C Ruminococcus bromii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
 recombinant enzyme Ruminococcus bromii
5
-
 substrate: soluble starch Ruminococcus bromii

pH Range

pH Minimum pH Maximum Comment Organism
4 9 30% of maxial activity at pH 4.0 and pH 9.0, profile overview Ruminococcus bromii
4 9 pH 4.0: about 30% of maximal activity, pH 9.0: about 30% of maximal activity, substrate: soluble starch Ruminococcus bromii

General Information

General Information Comment Organism
evolution the enzyme belongs to the glycosyl hydrolase family 13, GH13 Ruminococcus bromii
physiological function the enzyme may have a role to provide branched maltooligosaccharides to stimulate the growth of beneficial microorganisms in the human intestine Ruminococcus bromii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
15.4
-
 amylopectin 45°C, pH 5.0 Ruminococcus bromii
19.3
-
 pullulan 45°C, pH 5.0 Ruminococcus bromii
65.8
-
 starch 45°C, pH 5.0 Ruminococcus bromii
164
-
 amylose 45°C, pH 5.0 Ruminococcus bromii