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 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 | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Ruminococcus bromii | - |
- |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ruminococcus bromii | - |
- |
- |
Ruminococcus bromii | A0A2N0UQI0 | - |
- |
Ruminococcus bromii ATCC 27255 | - |
- |
- |
Ruminococcus bromii ATCC 27255 | A0A2N0UQI0 | - |
- |
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 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 | 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 | Comment | Organism |
---|---|---|
? | x * 60000, recombinant His6-tagged enzyme, SDS-PAGE, x * 58500, about, sequence calculation | Ruminococcus bromii |
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 Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
recombinant enzyme | Ruminococcus bromii |
45 | - |
substrate: soluble starch | Ruminococcus bromii |
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 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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
recombinant enzyme | Ruminococcus bromii |
5 | - |
substrate: soluble starch | Ruminococcus bromii |
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 | 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 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 |