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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.1 expression in Escherichia coli BL21 (DE3) Ruminococcus bromii
3.2.1.1 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
3.2.1.135 expression in Escherichia coli BL21 (DE3) Ruminococcus bromii

KM Value [mM]

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

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.1 extracellular
-
 Ruminococcus bromii
-
-
3.2.1.135 extracellular
-
 Ruminococcus bromii
-
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.135 pullulan + H2O Ruminococcus bromii the enzyme may have a role to provide branched maltooligosaccharides to stimulate the growth of beneficial microorganisms in the human intestine panose + ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.1 Ruminococcus bromii
-
-
-
3.2.1.1 Ruminococcus bromii A0A2N0UQI0
-
-
3.2.1.1 Ruminococcus bromii ATCC 27255
-
-
-
3.2.1.1 Ruminococcus bromii ATCC 27255 A0A2N0UQI0
-
-
3.2.1.135 Ruminococcus bromii
-
-
-
3.2.1.135 Ruminococcus bromii ATCC 27255
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.1
-
 Ruminococcus bromii
3.2.1.1 recombinant His6-tagged enzyme from Escherichia coli strain BL21-CodonPlus (DE3)-RP by nickel affinity chromatography and ultrafiltration Ruminococcus bromii
3.2.1.135
-
 Ruminococcus bromii

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.1 46.1
-
 purified recombinant His-tagged enzyme, pH 5.0, 45°C, substrate amylopectin Ruminococcus bromii
3.2.1.1 72.5
-
 45°C, pH 5.0 Ruminococcus bromii
3.2.1.1 72.5
-
 purified recombinant His-tagged enzyme, pH 5.0, 45°C, substrate pullulan Ruminococcus bromii
3.2.1.1 254
-
 purified recombinant His-tagged enzyme, pH 5.0, 45°C, substrate soluble starch Ruminococcus bromii
3.2.1.1 384
-
 purified recombinant His-tagged enzyme, pH 5.0, 45°C, substrate amylose Ruminococcus bromii
3.2.1.135 72.5
-
 45°C, pH 5.0 Ruminococcus bromii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.1 amylopectin + H2O at 12% of the activity as compared to amylose Ruminococcus bromii ?
-
 ?
3.2.1.1 amylopectin + H2O at 12% of the activity as compared to amylose Ruminococcus bromii ATCC 27255 ?
-
 ?
3.2.1.1 amylopectin + H2O 0.5% substrate solution, 12% activity compared to amylose Ruminococcus bromii maltose + D-glucose
-
 ?
3.2.1.1 amylose + H2O
-
 Ruminococcus bromii ?
-
 ?
3.2.1.1 amylose + H2O
-
 Ruminococcus bromii ATCC 27255 ?
-
 ?
3.2.1.1 amylose + H2O 0.5% substrate solution, best substrate Ruminococcus bromii maltose + D-glucose
-
 ?
3.2.1.1 amylose + H2O 0.5% substrate solution, best substrate Ruminococcus bromii ATCC 27255 maltose + D-glucose
-
 ?
3.2.1.1 gamma-cyclodextrin + H2O at 3.9% of the activity as compared to amylose Ruminococcus bromii ?
-
 ?
3.2.1.1 gamma-cyclodextrin + H2O at 3.9% of the activity as compared to amylose Ruminococcus bromii ATCC 27255 ?
-
 ?
3.2.1.1 maltopentaose + H2O
-
 Ruminococcus bromii maltose + ?
-
 ?
3.2.1.1 maltopentaose + H2O
-
 Ruminococcus bromii maltotriose + maltose + D-glucose
-
 ?
3.2.1.1 maltotetraose + H2O
-
 Ruminococcus bromii 2 maltose
-
 ?
3.2.1.1 maltotetraose + H2O
-
 Ruminococcus bromii ATCC 27255 2 maltose
-
 ?
3.2.1.1 maltotetraose + H2O
-
 Ruminococcus bromii maltose + ?
-
 ?
3.2.1.1 maltotriose + H2O
-
 Ruminococcus bromii maltose + D-glucose
-
 ?
3.2.1.1 maltotriose + H2O
-
 Ruminococcus bromii ATCC 27255 maltose + D-glucose
-
 ?
3.2.1.1 maltotriose + H2O
-
 Ruminococcus bromii maltose + ?
-
 ?
3.2.1.1 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 ?
-
 ?
3.2.1.1 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 ?
-
 ?
3.2.1.1 pullulan + H2O 0.5% substrate solution, 18.9% activity compared to amylose Ruminococcus bromii panose + ?
-
 ?
3.2.1.1 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 + ?
-
 ?
3.2.1.1 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 + ?
-
 ?
3.2.1.1 pullulan + H2O 0.5% substrate solution, 18.9% activity compared to amylose Ruminococcus bromii ATCC 27255 panose + ?
-
 ?
3.2.1.1 soluble starch + H2O 0.5% substrate solution, 66.2% activity compared to amylose Ruminococcus bromii maltose + D-glucose
-
 ?
3.2.1.1 starch + H2O at 66.2% of the activity as compared to amylose Ruminococcus bromii ?
-
 ?
3.2.1.1 starch + H2O at 66.2% of the activity as compared to amylose Ruminococcus bromii ATCC 27255 ?
-
 ?
3.2.1.135 pullulan + H2O the enzyme may have a role to provide branched maltooligosaccharides to stimulate the growth of beneficial microorganisms in the human intestine Ruminococcus bromii panose + ?
-
 ?
3.2.1.135 pullulan + H2O the enzyme is an alpha-amylase with neopullulanase-like activity Ruminococcus bromii panose + ?
-
 ?

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
3.2.1.1 amy5
-
 Ruminococcus bromii
3.2.1.1 RB5AMG_01539
-
 Ruminococcus bromii
3.2.1.1 Rbamy5
-
 Ruminococcus bromii
3.2.1.1 Ruminococcus bromii intermediary alpha-amylase 5
-
 Ruminococcus bromii
3.2.1.135 Rbamy5
-
 Ruminococcus bromii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.1 45
-
 recombinant enzyme Ruminococcus bromii
3.2.1.1 45
-
 substrate: soluble starch Ruminococcus bromii

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.1 30 50 30°C: about 40% of maximal activity, 50°C: about 40% of maximal activity, substrate: soluble starch Ruminococcus bromii
3.2.1.1 30 50 40% of maximal activity at 30°C and 50°C, profile overview Ruminococcus bromii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.1 35
-
 140 min, no loss of activity Ruminococcus bromii
3.2.1.1 35
-
 purified recombinant enzyme, completely stable for 140 min Ruminococcus bromii
3.2.1.1 40
-
 120 min, about 45% loss of activity Ruminococcus bromii
3.2.1.1 40
-
 purified recombinant enzyme, loss of 50% activity after 140 min Ruminococcus bromii
3.2.1.1 45
-
 20 min, about 70% loss of activity Ruminococcus bromii
3.2.1.1 45
-
 purified recombinant enzyme, inactivation within about 27 min Ruminococcus bromii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.1 137
-
 amylopectin 45°C, pH 5.0 Ruminococcus bromii
3.2.1.1 137
-
 amylopectin recombinant His-tagged enzyme, pH 5.0, 45°C Ruminococcus bromii
3.2.1.1 297
-
 pullulan 45°C, pH 5.0 Ruminococcus bromii
3.2.1.1 297
-
 pullulan recombinant His-tagged enzyme, pH 5.0, 45°C Ruminococcus bromii
3.2.1.1 1014
-
 amylose 45°C, pH 5.0 Ruminococcus bromii
3.2.1.1 1014
-
 amylose recombinant His-tagged enzyme, pH 5.0, 45°C Ruminococcus bromii
3.2.1.1 1055
-
 starch 45°C, pH 5.0 Ruminococcus bromii
3.2.1.1 1055
-
 soluble starch recombinant His-tagged enzyme, pH 5.0, 45°C Ruminococcus bromii
3.2.1.135 297
-
 pullulan 45°C, pH 5.0 Ruminococcus bromii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.1 5
-
 recombinant enzyme Ruminococcus bromii
3.2.1.1 5
-
 substrate: soluble starch Ruminococcus bromii

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.1 4 9 30% of maxial activity at pH 4.0 and pH 9.0, profile overview Ruminococcus bromii
3.2.1.1 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

EC Number General Information Comment Organism
3.2.1.1 evolution the enzyme belongs to the glycosyl hydrolase family 13, GH13 Ruminococcus bromii
3.2.1.1 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
3.2.1.135 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.1 15.4
-
 amylopectin 45°C, pH 5.0 Ruminococcus bromii
3.2.1.1 19.3
-
 pullulan 45°C, pH 5.0 Ruminococcus bromii
3.2.1.1 65.8
-
 starch 45°C, pH 5.0 Ruminococcus bromii
3.2.1.1 164
-
 amylose 45°C, pH 5.0 Ruminococcus bromii
3.2.1.135 19.3
-
 pullulan 45°C, pH 5.0 Ruminococcus bromii