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BGALD-RO

Roche

β-Galactose Dehydrogenase

from Pseudomonas fluorescens

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Synonym(s):
D-galactose dehydrogenase, D-galactose:NAD+ 1-oxidoreductase
Enzyme Commission number:
1.1.1.48 (BRENDA, IUBMB)
UNSPSC Code:
12352204

biological source

Pseudomonas fluorescens

Quality Level

100

recombinant

expressed in E. coli

form

suspension

specific activity

~5 units/mg protein (At 25 °C with D-galactose as the substrate; standardized with BSA.)

mol wt

64,000 Da

packaging

pkg of 1 mL (10104973001 [1 mg])
pkg of 1 mL (10104981001 [5 mg])

manufacturer/tradename

Roche

technique(s)

activity assay: suitable

color

white

optimum pH

8.4 (0.1 M in Tris buffer)

pH range

4.6-9.2

solubility

water: soluble

suitability

suitable for enzyme test

UniProt accession no.

 (T1WEA6 · T1WEA6_PSEFL)

foreign activity

ADH <0.01%
LDH <0.5%
NADH oxidase <0.1%
galactosidase <0.01%

storage temp.

2-8°C

Related Categories

General description

β-Galactose Dehydrogenase plays a crucial role in galactose metabolism by catalyzing the pyridine-nucleotide-dependent dehydrogenation of β-D-galactopyranose to yield NADH and D-galactono-l,5-1actone. D-galactose dehydrogenase from P. fluorescens is a member of the oxidoreductases family. It is composed of two identical subunits and has a molecular weight of 64,000. Additionally, the enzyme contains two binding sites.

Specificity

Galactose dehydrogenase will oxidize D-galactose [Km = 0.7 mM (30 °C, pH 9.1); relative rate = 1.00], L-arabinose (relative rate = 0.47), D-fucose (relative rate = 1.05), 2-deoxy-D-galactose (relative rate = 0.27), 2-amino-2-deoxy-D-galactose [galactosamine], (relative rate = 0.02; pH 8.6).
Galactose dehydrogenase does not oxidize L-galactose, D-arabinose, D-glucose, L-fucose, D-ribose, D-xylose, D-glucuronic acid, D-galacturonic acid or D-galactose-6-phosphate. The enzyme will use either NAD (Km = 0.24 mM; relative rate = 1.0) or NADP (Km = 2.3 mM; relative rate = 0.32).

Application

β-Galactose Dehydrogenase has been used as a reporter enzyme in neuraminidase (NA) kinetic parameters for the natural substrates 6′-Sialyl-N-acetyllactosamine (6SLN) and 3′-Sialyl-N-acetyllactosamine (3SLN).

Quality

Contaminants: <0.01% ADH, <0.01% ?-galactosidase, <0.1% NADH oxidase, <0.5% LDH

Sequence

Galactose dehydrogenase from Pseudomonas fluorescens (MW 64,000 D) is a dimer with subunits of equal size (subunit MW = 32,000 D).

Unit Definition

One unit galactose dehydrogenase will oxidize 1 mol of β-D-galactose in one minute at +25 °C and pH 8.6.
The QC assay produces 1 mol of NADH per mol of galactose oxidized.

Physical form

Suspension in 3.2 M ammonium sulfate solution, 1 mM EDTA, pH approximately 6

Other Notes

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

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

No data available

Flash Point(C)

No data available

Regulatory Information

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[29] d-Galactose dehydrogenase from Pseudomonas fluorescens
E. Maier and G. Kurz
Methods in Enzymology, 89.0, 176-181 (1982)
Biophysical Measurement of the Balance of Influenza A Hemagglutinin and Neuraminidase Activities*
Benton, et al.
The Journal of Biological Chemistry, 290, 6516-6521 (2015)
Biophysical measurement of the balance of influenza a hemagglutinin and neuraminidase activities
Benton D, et al.
The Journal of biological chemistry, 290(10), 6516-6521 (2015)
Recombinant D-galactose dehydrogenase partitioning in aqueous two-phase systems: effect of pH and concentration of PEG and ammonium sulfate
Kianmehr A, et al.
Bioresources and bioprocessing, 1.0 (2014)
Modelling severe Staphylococcus aureus sepsis in conscious pigs: are implications for animal welfare justified?
Olsen, et al.
BMC Research Notes, 9 (2016)

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