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Literature summary for 3.2.1.23 extracted from
- Purification and characterization of a novel beta-galactosidase with transglycosylation activity from Bacillus megaterium 2-37-4-1 (2008), Appl. Biochem. Biotechnol., 158, 192-199.
Application
| Application | Comment | Organism |
|---|---|---|
| nutrition | wide acceptor specificity for transglycosylation reactions capable of the synthesis of important galactosyl compounds such as alkyl glycoside. The enzyme may be a novel tool for enzymatic synthesis with applications in the food, healthcare, and pharmaceutical industries | Priestia megaterium |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| cloned in pET15b vector, overexpression in Escherichia coli BL21 (DE3) pLysS | Priestia megaterium |
| expression in Escherichia coli | Priestia megaterium |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 9.5 | - |
o-nitrophenyl-beta-D-galactoside | - |
Priestia megaterium |  |
| 9.5 | - |
2-nitrophenyl-beta-D-galactoside | 50°C | Priestia megaterium | |
| 12.6 | - |
lactose | - |
Priestia megaterium | |
| 12.6 | - |
lactose | 50°C | Priestia megaterium | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 118000 | - |
1 * 118000, calculated, 1 * 120000, SDS-PAGE | Priestia megaterium |
| 120000 | - |
PAGE | Priestia megaterium |
| 120000 | - |
SDS-PAGE and native PAGE, enzyme is purified about 10.2fold by a five-step procedure | Priestia megaterium |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Priestia megaterium | A3FFK4 | - |
- |
| Priestia megaterium | A3FFK4 | expression in Escherichia coli | - |
| Priestia megaterium 2-37-4-1 | A3FFK4 | - |
- |
| Priestia megaterium 2-37-4-1 | A3FFK4 | expression in Escherichia coli | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Priestia megaterium |
| ammonium sulfate precipitation, followed by desalting, and sequentially applied to a DEAE Sepharose Fast Flow column, a Gigapite K-100 S column and a para-aminophenyl-1-thio-beta-D-galactopyranoside (APTG) affinity chromatography column | Priestia megaterium |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 59.9 | - |
- |
Priestia megaterium |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 4°C, pH 6.0-9.0, stable for 24 h | Priestia megaterium |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-nitrophenyl beta-D-galactoside + H2O | - |
Priestia megaterium | 2-nitrophenol + D-galactose | - |
? | |
| 2-nitrophenyl beta-D-galactoside + H2O | - |
Priestia megaterium 2-37-4-1 | 2-nitrophenol + D-galactose | - |
? | |
| 4-nitrophenyl beta-D-galactoside + H2O | 89% of the activity with 2-nitrophenyl-beta-D-galactoside | Priestia megaterium | 4-nitrophenol + D-galactose | - |
? | |
| 4-nitrophenyl beta-D-galactoside + H2O | 89% of the activity with 2-nitrophenyl-beta-D-galactoside | Priestia megaterium 2-37-4-1 | 4-nitrophenol + D-galactose | - |
? | |
| lactose + H2O | - |
Priestia megaterium | D-glucose + D-galactose | - |
? | |
| lactose + H2O | - |
Priestia megaterium | D-galactose + D-glucose | - |
? | |
| additional information | highest activity for o-nitrophenyl-beta-D-galactoside and 89% activity using p-nitrophenyl-beta-D-galactopyranoside as the substrate, no activity for p-nitrophenyl-alpha-D-galactoside, o-nitrophenyl-alpha-D-galactoside, o-nitrophenyl-alpha-D-glucoside, o-nitrophenyl-beta-D-glucoside, p-nitrophenyl-N-acetyl-beta-D-galactosaminide, p-nitrophenyl-N-acetyl-alpha-D-galactosaminide | Priestia megaterium | ? | - |
? | |
| additional information | wide acceptor specificity for transglycosylation reactions capable of the synthesis of important galactosyl compounds such as alkyl glycoside, beta-galactosidase prefers fructose, glucose, sorbitose, sucrose, cellobiose, sorbitol, mannitol, and alkyl alcohol as acceptor than other saccharides | Priestia megaterium | ? | - |
? | |
| additional information | enzyme additionally has transglycosylation activity with a series of acceptors, including pentose, hexose, hydroxyl, and alkyl alcohol using 2-nitrophenyl-beta-D-galactoside as a donor | Priestia megaterium | ? | - |
? | |
| additional information | highest activity for o-nitrophenyl-beta-D-galactoside and 89% activity using p-nitrophenyl-beta-D-galactopyranoside as the substrate, no activity for p-nitrophenyl-alpha-D-galactoside, o-nitrophenyl-alpha-D-galactoside, o-nitrophenyl-alpha-D-glucoside, o-nitrophenyl-beta-D-glucoside, p-nitrophenyl-N-acetyl-beta-D-galactosaminide, p-nitrophenyl-N-acetyl-alpha-D-galactosaminide | Priestia megaterium 2-37-4-1 | ? | - |
? | |
| additional information | wide acceptor specificity for transglycosylation reactions capable of the synthesis of important galactosyl compounds such as alkyl glycoside, beta-galactosidase prefers fructose, glucose, sorbitose, sucrose, cellobiose, sorbitol, mannitol, and alkyl alcohol as acceptor than other saccharides | Priestia megaterium 2-37-4-1 | ? | - |
? | |
| additional information | enzyme additionally has transglycosylation activity with a series of acceptors, including pentose, hexose, hydroxyl, and alkyl alcohol using 2-nitrophenyl-beta-D-galactoside as a donor | Priestia megaterium 2-37-4-1 | ? | - |
? | |
| o-nitrophenyl-beta-D-galactoside + H2O | - |
Priestia megaterium | o-nitrophenol + beta-D-galactose | - |
? | |
| p-nitrophenyl-beta-D-galactopyranoside + H2O | - |
Priestia megaterium | p-nitrophenol + beta-D-galactose | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1* 120000 SDS-PAGE | Priestia megaterium |
| monomer | 1 * 118000, calculated, 1 * 120000, SDS-PAGE | Priestia megaterium |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| beta-galactosidase | - |
Priestia megaterium |
| BgaBM | - |
Priestia megaterium |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
- |
Priestia megaterium |
| 55 | - |
by incubating the enzyme with o-nitrophenyl-beta-D-galactoside for 10 min at temperatures ranging from 0°C to 70°C | Priestia megaterium |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
favorable thermostability within a broad range of pHs (6.09.0), its activity decreases below 10% for 15 min at 60°C without substrate protection | Priestia megaterium |
| 40 | - |
60 min, 40% loss of activity | Priestia megaterium |
| 50 | - |
60 min, 50% loss of activity | Priestia megaterium |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | 8 | - |
Priestia megaterium |
| 7.5 | 8 | at 55°C by assaying the enzymatic activity at pH values ranging from 3.0 to 10.0 | Priestia megaterium |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 9 | below 40°C | Priestia megaterium |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 9 | - |
Priestia megaterium |
| 6 | 9 | by incubating the enzyme in the presence of different pH buffers at 4°C for 24 h | Priestia megaterium |
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