10105031001

β-Galactosidase

from E. coli overproducer

• Synonym(s): β-galactosidase
• Enzyme Commission number: 3.2.1.23 (BRENDA, IUBMB)
• UNSPSC Code: 12352204

Properties

• biological source: Escherichia coli
• Quality Level: 100
• form: suspension
• mol wt: 540 kDa
• packaging: pkg of 1,500 U
• manufacturer/tradename: Roche
• concentration: 5.00 mg/mL
• technique(s): activity assay: suitable
• optimum pH: 7
• suitability: suitable for UV spectrophotometry and general use
• application(s): life science and biopharma
• shipped in: wet ice
• storage temp.: 2-8°C

Description

General description

β-Galactosidase or β-D-Galactoside galactohydrolase is basically a tetramer made up of identical four polypeptide chains, each constituting 1023 amino acids. It is very specific for D-galactose and requires K⁺ or Na⁺ and Mg²⁺ to be fully active. β-Galactosidase enzyme with an oxygen glycosidic bond catalyzes reactions with β-d-galactopyranosides.[1]

At 37 °C with 2-nitrophenyl-β-D-galactoside as the substrate, approximately 30 U/mg at 25 °C with lactose as the substrate; standardized with BSA.

Specificity

Cleaves terminal galactose residues that are β1,4-linked to a monosaccharide, oligosaccharide, or glycopeptide.

Application

β-galactosidase has been used in enzyme-linked immunosorbent assay (ELISA).[2] Use β-Galactosidase to produce a calibration curve in enzymatic assays.

Quality

Contaminants: <0.01% GIDH, GPT, LDH, MDH, and oxaloacetate decarboxylase, each

Physical form

Suspension, in 3.2 M ammonium sulfate solution, pH approximately 6, crystalline

Preparation Note

The ammonium sulfate preparation is stable at 2 to 8 °C until the expiration date printed on the label. Use directly for most applications, e.g., quantitation of lactose.

In the absence of ammonium sulfate, solutions of β-galactosidase should be stabilized with Mg²⁺ (89 mM) and a thiol reagent (1mM β-mercaptoethanol, 1 mM dithiothreitol) [Beutler, 1984]. The thiol slows the formation of enzyme dimers resulting from intramolecular disulfide bridges.

Activator: K (50 mM) is required for activation (lactose hydrolysis).
Na (50 mM) is also an activator, particularly for hydrolysis of 2-nitrophenyl-β-D-galactopyranoside.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): No data available
• Flash Point(C): No data available