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Literature summary extracted from

  • Li, Y.; Wang, H.; Lu, L.; Li, Z.; Xu, X.; Xiao, M.
    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.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.2.1.23 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)

EC Number Cloned (Comment) Organism
3.2.1.23 cloned in pET15b vector, overexpression in Escherichia coli BL21 (DE3) pLysS Priestia megaterium
3.2.1.23 expression in Escherichia coli Priestia megaterium

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.23 9.5
-
 o-nitrophenyl-beta-D-galactoside
-
 Priestia megaterium
3.2.1.23 9.5
-
 2-nitrophenyl-beta-D-galactoside 50°C Priestia megaterium
3.2.1.23 12.6
-
 lactose
-
 Priestia megaterium
3.2.1.23 12.6
-
 lactose 50°C Priestia megaterium

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.23 118000
-
 1 * 118000, calculated, 1 * 120000, SDS-PAGE Priestia megaterium
3.2.1.23 120000
-
 PAGE Priestia megaterium
3.2.1.23 120000
-
 SDS-PAGE and native PAGE, enzyme is purified about 10.2fold by a five-step procedure Priestia megaterium

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.23 Priestia megaterium A3FFK4
-
-
3.2.1.23 Priestia megaterium A3FFK4 expression in Escherichia coli
-
3.2.1.23 Priestia megaterium 2-37-4-1 A3FFK4
-
-
3.2.1.23 Priestia megaterium 2-37-4-1 A3FFK4 expression in Escherichia coli
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.23
-
 Priestia megaterium
3.2.1.23 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]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.23 59.9
-
-
 Priestia megaterium

Storage Stability

EC Number Storage Stability Organism
3.2.1.23 4°C, pH 6.0-9.0, stable for 24 h Priestia megaterium

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.23 2-nitrophenyl beta-D-galactoside + H2O
-
 Priestia megaterium 2-nitrophenol + D-galactose
-
 ?
3.2.1.23 2-nitrophenyl beta-D-galactoside + H2O
-
 Priestia megaterium 2-37-4-1 2-nitrophenol + D-galactose
-
 ?
3.2.1.23 4-nitrophenyl beta-D-galactoside + H2O 89% of the activity with 2-nitrophenyl-beta-D-galactoside Priestia megaterium 4-nitrophenol + D-galactose
-
 ?
3.2.1.23 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
-
 ?
3.2.1.23 lactose + H2O
-
 Priestia megaterium D-glucose + D-galactose
-
 ?
3.2.1.23 lactose + H2O
-
 Priestia megaterium D-galactose + D-glucose
-
 ?
3.2.1.23 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 ?
-
 ?
3.2.1.23 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 ?
-
 ?
3.2.1.23 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 ?
-
 ?
3.2.1.23 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 ?
-
 ?
3.2.1.23 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 ?
-
 ?
3.2.1.23 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 ?
-
 ?
3.2.1.23 o-nitrophenyl-beta-D-galactoside + H2O
-
 Priestia megaterium o-nitrophenol + beta-D-galactose
-
 ?
3.2.1.23 p-nitrophenyl-beta-D-galactopyranoside + H2O
-
 Priestia megaterium p-nitrophenol + beta-D-galactose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.23 monomer 1* 120000 SDS-PAGE Priestia megaterium
3.2.1.23 monomer 1 * 118000, calculated, 1 * 120000, SDS-PAGE Priestia megaterium

Synonyms

EC Number Synonyms Comment Organism
3.2.1.23 beta-galactosidase
-
 Priestia megaterium
3.2.1.23 BgaBM
-
 Priestia megaterium

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.23 55
-
-
 Priestia megaterium
3.2.1.23 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]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.23 additional information
-
 favorable thermostability within a broad range of pHs (6.0–9.0), its activity decreases below 10% for 15 min at 60°C without substrate protection Priestia megaterium
3.2.1.23 40
-
 60 min, 40% loss of activity Priestia megaterium
3.2.1.23 50
-
 60 min, 50% loss of activity Priestia megaterium

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.23 7.5 8
-
 Priestia megaterium
3.2.1.23 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

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.23 6 9 below 40°C Priestia megaterium

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.23 6 9
-
 Priestia megaterium
3.2.1.23 6 9 by incubating the enzyme in the presence of different pH buffers at 4°C for 24 h Priestia megaterium