Nacalai USA, Inc. - Products

3α-Hydroxysteroid: NAD(P)⁺ Oxidoreductase [EC 1.1.1.50]

Origin

Pseudomonas testosteroni

Reaction

3Α-hydroxysteroid + NAD(P)⁺

3-Oxosteroid + NAD(P)H+H⁺

Appearance

White amorphous powder.

Activity

More than 90 units/mg protein.

Unit definition

One unit is the amount of enzyme required to oxidize 1 ?mol of Androsterone as a substrate per minute in the presence of NAD at pH 8.9 at 25°C.

Storage

Stable for one year when stored below 5°C and also stable at room temperature for at least one week. For prolonged storage, keep at -20°C.

Contaminants

Malate Dehydrogenase < 0.01%
Lactate Dehydrogenase < 0.01%
Alcohol Dehydrogenase < 0.01%
β-Hydroxysteroid dehydrogenase < 0.5 %

Properties

Molecular weight	---	Approx. 37,000 Da
Optimum pH	---	10.2 ~ 10.5
Optimum temperature	---	50°C
pH stability	---	6.0 ~ 9.5 (30°C, 17 hr)
Thermal stability	---	below 50°C (pH 7.2, 10 min)
Michaelis constant	---	6.7x10-⁶ M (Androsterone) 8.3x10-⁶ M (Na-cholate) 6.7x10-⁵ M (NAD)

Uses

Determination of bile acid

Ordering information

Product	Storage	Cat.No.	Quantity	Price(US$)
3α-Hydroxysteroid dehydrogenase 3α-Hydroxysteroid : NAD(P)⁺ oxidoreductase [EC 1.1.1.50]	5°C	18949-34	10 units	121
3α-Hydroxysteroid dehydrogenase 3α-Hydroxysteroid : NAD(P)⁺ oxidoreductase [EC 1.1.1.50]	5°C	18949-76	50 units	286

3(or 17)β-Hydroxysteroid:NAD(P)⁺ Oxidoreductase [EC 1.1.1.51]

Origin

Pseudomonas testosteroni

Reaction

(1) epi-androsterone + NAD(P)⁺

5α-Androstene-3,17-dione + NAD(P)H+H⁺
(2) testosterone + NAD(P)⁺

4-Androstene-3,17-dione + NAD(P)H+H⁺

Appearance

White amorphous powder.

Activity

More than 25 units/mg protein.

Unit definition

One unit oxidize 1 µmol of testosterone as a substrate per minute in the presence of NAD at pH 8.9 at 25°C.

Contaminants

Malate Dehydrogenase < 0.01%
Lactate Dehydrogenase < 0.01%
Alcohol Dehydrogenase < 0.01%
3α-Hydroxysteroid Dehydrogenase approx. 0.1 %

Storage

Stable for one year when stored below 5°C and also stable at room temperature for at least one week. For prolonged storage, keep at -20°C.

Properties

Molecular weight	---	Approx. 80,000 Da
Optimum pH	---	10.1 ~ 10.3
Optimum temperature	---	34°C
pH stability	---	7.0 ~ 8.7
Thermal stability	---	below 42°C (pH 7.2, 10 min)
Michaelis constant	---	2.5x10-⁶ M (Testosterone) 1.8x10-⁴ M (NAD)

Ordering information

Product	Storage	Cat.No.	Quantity	Price(US$)
β-Hydroxysteroid dehydrogenase 3(or17) β-Hydroxysteroid : NAD(P)⁺ oxidoreductase [EC 1.1.1.51]	5°C	18950-94	10 units	97
β-Hydroxysteroid dehydrogenase 3(or17) β-Hydroxysteroid : NAD(P)⁺ oxidoreductase [EC 1.1.1.51]	5°C	18950-36	50 units	269

Origin

Bacillus licheniformis

Reaction

NADH + H⁺ + O₂

NAD⁺ + H₂O₂

Appearance

Yellow amorphous powder.

Activity

More than 50 units/mg protein.

Unit definition

One unit is the amount of enzyme required to oxidize 1 µmol of NADH per minute at pH 7.0 at 30°C.

Contaminants

Sometimes, trace amount of catalase might be detected. Therefore, the addition of 10 mM NaN₃ into the reaction mixture is recommended when the complete elimination of catalase is needed.

Storage

Stable for one year when stored below 5°C and also stable at room temperature for at least one week. For prolonged storage, keep at -20°C.

Properties

Molecular weight	---	Approx. 240,000 Da
Optimum pH	---	6.5 ~ 7.5
Optimum temperature	---	45°C
pH stability	---	7.0 ~ 8.5
Thermal stability	---	below 30°C (pH 7.5, 10 min)and below 40°C (in the coexistence of 0.1% bovine serum albumin,pH 7.5, 10 min)
Michaelis constant	---	3.2x10-⁵ M ( NADH ) 6.7x10-⁶ M ( FAD )
Substrate specificity	---	In the absence of added FAD both NADH and NADPH are oxidized equally, but by the addition of FAD ( about 30 µM ) to reaction mixture the reaction velocity to NADH is accelerated about 20 ~ 30 times in contrast to 2 ~ 3 times of NADPH. Accordingly, the substrate specificity of NADH is about 10 times larger than that of NADPH in the presence of added FAD.

Ordering information

Product	Storage	Cat.No.	Quantity	Price(US$)
NADH oxidase	5°C	23626-94	5 units	120
NADH oxidase	5°C	23626-52	25 units	480

Origin

Escherichia coli

Reaction

N-Acetylneuraminate

N-Acetyl-D-mannosamine + Pyruvate

Appearance

White amorphous powder.

Activity

More than 30 units/mg protein.

Unit definition

One unit is the amount of enzyme required to liberates 1 µmol of N-Acetylmannosamine (or Pyruvic Acid) per minute at pH 7.7 at 37°C, using N-Acetylneuraminic Acid (NANA) as a substrate.

Storage

Stable for one year when stored below 5°C. For prolonged storage, keep at -20°C.

Contaminant

Free from NADH oxidase

Properties

Molecular weight	---	Approx. 98,000 Da (gel filtration)
Optimum pH	---	7.5 ~ 8.0
pH stability	---	6.0 ~ 9.0
Thermal stability	---	below 65°C (pH 7.0, 20 min)
Substrate specificity	---	N-Glycolylneuraminic Acid (NGNA) is cleaved as well as NANA. Km = 3.6 mM (NANA), 4.3 mM (NGNA)

Uses

Enzymatic determination of Sialic Acid and enzymatic syntheses of novel Sialic Acid derivatives.

Ordering information

Product	Storage	Cat.No.	Quantity	Price(US$)
N-Acetylneuraminic acid aldolase N-Acetylneuraminate pyruvate lyase [EC 4.1.3.3]	5°C	00628-84	10 units	57

Origin

Escherichia coli JM109 carrying the plasmid encoding Acylglucosamine 2-Epimerase (AGE) gene from hog kidney.

Reaction

N-Acetyl-D-glucosamine

N-Acetyl-D-mannosamine. AGE can tightly bind to renin and inhibit the renin activity.

Appearance

Lyophilized powder.

Activity

More than 30 units/mg protein.

Unit definition

One unit of enzyme activity is the quantity that produced 1 µmol of N-Acetyl-D-glucosamine per minute at 25°C at pH 8.9 using N-Acetyl-Dmannosamine as a substrate.

Storage

Stable for one year when stored below 5°C. For prolonged storage, keep at -20°C.

Properties

Molecular weight	---	Approx. 93,000 Da (sedimentation equilibrium)
Subunit	---	46,600 (SDS-PAGE)
Optimum pH	---	6.8
Optimum temperature	---	47°C
pH stability	---	6.0 ~ 8.0
Thermal stability	---	below 37°C
Activator	---	ATP (or dATP) and MgCl₂
Substrate specificity	---	N-Acetyl-D-glucosamine and N-Acetyl-D-mannosamine
Michaelis constant	---	7.4x10-⁶ M (N-Acetyl-D-glucosamine) 6.3x10-⁶ M (N-Acetyl-D-mannosamine) 1.8x10-⁷ M (ATP)

Ordering information

Product	Storage	Cat.No.	Quantity	Price(US$)
Acylglucosamine 2-epimerase (Renin binding protein) Acylglucosamine 2-epimerase [EC 5.1.3.8]	5°C	00853-11	1 unit	230
Acylglucosamine 2-epimerase (Renin binding protein) Acylglucosamine 2-epimerase [EC 5.1.3.8]	5°C	00853-24	5 units	690

Origin

Arthrobacter ureafaciens

Reaction

Sialyl compound

Sialic Acid + Asialocompound

Appearance

White amorphous powder.

Activity

More than 80 units/mg protein for N-Acetylneuraminyllactose (NANA-lactose).

Unit definition

One unit is the amount of enzyme required to liberate 1 µmol of N-Acetylneuraminic Acid (NANA) per minute at pH 5.0 at 37°C.

Storage

Stable for one year when stored below 5°C. For prolonged storage, keep at -20°C.

Contaminant

Enzyme activities mentioned below cannot be detected. Protease , N-Acetylneuraminic Acid Aldolase, Glycosidase such as α-Glucosidase, β-Glucosidase, α-Galactosidase, β-Galactosidase, α-Mannosidase, α-Fucosidase, N-Acetyl-α-glucosaminidase, N-Acetyl-β-glucosaminidase, N-Acetyl-agalactosaminidase, N-Acetyl-β-galactosaminidase, N-Acetyl-α-mannosaminidase and N-Acetyl-β-mannosaminidase.

Properties

Molecular weight	---	Approx. 52,000 Da, 66,000 Da and 88,000 Da (gel filtration, SDS-PAGE)
Optimum pH	---	4.5 ~ 5.5 (NANA-lactose as a substrate)
pH stability	---	4.5 ~ 9.5
Thermal stability	---	below 60°C (pH 5.0, 20 min)
Substrate specificity	---	The α-ketosidic linkage of N-Glycolylneuraminic Acid (NGNA) can be hydrolyzed as well as that of NANA. This enzyme cleaves α(2 3), α(2 6) and α(2 8) linkages of N-Acetylneuraminic Acid in glycoconjugates. The acivity is independent on Ca²⁺ and is not inhibited by EDTA, which is in striking contrast to Vibrio cholerae Neuraminidase, and is not or slightly inhibited by inhibitors such as Monoiodoacetate, p-Chloromercuribenzoate and HgCl₂, which is in striking contrast to Clostridium perfringens Neuraminidase.

Ordering information

Product	Storage	Cat.No.	Quantity	Price(US$)
Neuraminidase [Sialidase] Acylneuraminyl hydrolase [EC 3.2.1.18]	5°C	24229-61	1 unit	70
Neuraminidase [Sialidase] Acylneuraminyl hydrolase [EC 3.2.1.18]	5°C	24229-74	5 units	240

Origin

Arthrobacter ureafaciens

Appearance

White amorphous powder.

Activity

More than 80 units/mg protein for N-Acetylneuraminyllactose (NANA-lactose).
More than 20 units/mg protein for bovine submaxillary mucin. [at pH 5.0]
More than 40 units/mg protein for Colominic Acid. [at pH 4.5]
More than 60 units/mg protein for bovine brain ganglioside. [at pH 4.0]

Unit definition

One unit is the amount of enzyme required to liberate 1 µmol of N-Acetylneuraminic Acid (NANA) per minute at pH 5.0 at 37°C, using N-Acetylneuraminyllactose as a substrate.

Storage

Stable for one year when stored below 5°C. For prolonged storage, keep at -20°C.

Contaminant

Enzyme activities mentioned below cannot be detected. Protease , N-Acetylneuraminic Acid Aldolase, Glycosidase such as α-Glucosidase, β-Glucosidase, α-Galactosidase, β-Galactosidase, α-Mannosidase, α-Fucosidase, N-Acetyl-α-glucosaminidase, N-Acetyl-β-glucosaminidase, N-Acetyl-α-galactosaminidase, N-Acetyl-β-galactosaminidase, N-Acetyl-α-mannosaminidase and N-Acetyl-β-mannosaminidase.

Properties

Molecular weight	---	Approx. 52,000 Da (gel filtration, SDS-PAGE)
Optimum pH	---	3.8 ~ 4.4 ( for bovine brain ganglioside )
pH stability	---	3.5 ~ 10.0
Thermal stability	---	below 60°C (pH 5.0, 20 min)
Substrate specificity	---	Isozyme S cleaves α(2 3), α(2 6) and α(2 8) linkages of N-Acetylneuraminic Acid in glycoconjugates. In the absence of detergents and calcium ion, the enzyme hydrolyzes N-Acetylneuraminosyl moiety of polysialogangliosides and produces monosialoganglioside G_M1, while in the presence of detergents ( Na-cholate, Triton X-100 etc. ), G_M1 is further desialylated to asialoganglioside G_A1. the character of the enzyme is similar to Vibrio cholerae Neuraminidase except requirement of Ca²⁺ in the latter.

Ordering information

Product	Storage	Cat.No.	Quantity	Price(US$)
Neuraminidase isozyme S [Reagent for studies of gangliosides]	5°C	24238-41	1 unit	110
Neuraminidase isozyme S [Reagent for studies of gangliosides]	5°C	24238-54	5 units	420

Enzymes | Biochemical

3α-Hydroxysteroid Dehydrogenase

β-Hydroxysteroid Dehydrogenase

NADH Oxidase

N-Acetylneuraminic Acid Aldolase [EC 4.1.3.3]

Acylglucosamine 2-Epimerase (Renin Binding Protein) [EC 5.1.3.8]

Neuraminidase [Sialidase] [EC 3.2.1.18]

Neuraminidase Isozyme S