Temperature and pH optimum

The optimum pH and temperature are 5.5 and 25 °C, respectively. NADH peroxidase Specific activity (EC 1.11.1.1), Streptococcus 50 U/mg, 500 U/ml. faecalis ATCC 11700 Unit definition One unit is defined as the amount of enzyme required to + produce 1 mol of NAD from NADH in a reaction mixture Catalogue number: AE00201, 500 U (10 mg) containing 50 mM potassium phosphate buffer, pH 5.5, and 2 mM NADH, at 25 °C.

Protein sequence

Description MKVIVLGSSHGGYEAVEELLNLHPDAEIQWYEKGDFISFLSCGMQL NADH peroxidase (EC 1.11.1.1), systematically designated by YLEGKVKDVNSVRYMTGEKMESRGVNVFSNTEITAIQPKEHQVTV NADH:hydrogen-peroxide oxidoreductase, is a enzyme that KDLVSGEERVENYDKLIISPGAVPFELDIPGKDLDNIYLMRGRQWAI catalyzes the reversible conversion of NADH and hydrogen KLKQKTVDPEVNNVVVIGSGYIGIEAAEAFAKAGKKVTVIDILDRPL peroxide into NAD+ and water. It is a flavoprotein, thus GVYLDKEFTDVLTEEMEANNITIATGETVERYEGDGRVQKIVTDKN requiring the FAD as cofactor. The Nzytech NADH peroxidase AYDADLVVVAVGVRPNTAWLKGTLELHPNGLIKTDEYMRTSEPD is the Streptococcus faecalis enzyme expressed in Escherichia coli. The crystal structure of NADH peroxidase from VFAVGDATLIKYNPADTEVNIALATNARKQGRFAVKNLEEPVKPFP Streptococcus faecalis has been solved (PDB accession GVQGSSGLAVFDYKFASTGINEVMAQKLGKETKAVTVVEDYLMD number 1NPX). The enzyme is a symmetrical tetramer with a FNPDKQKAWFKLVYDPETTQILGAQLMSKADLTANINAISLAIQA fold similar to those of disulfide oxidoreductases. Swirl the KMTIEDLAYADFFFQPAFDKPWNIINTAALEAVKQER enzyme mix immediately prior to use. Enquire [email protected] to obtain any NADH peroxidase Purity mutant derivative.

NADH peroxidase has been determined to be >95% pure, according to SDS polyacrylamide gel electrophoresis (PAGE) References followed by Coomassie blue staining (Figure 1). Ross R.P. and Claiborne A. (1991) Journal of Molecular Biology 221, 857-871. kDa 96 Stehle et al. (1991) Journal of Molecular Biology 221, 1325– 66 1344. 48 40 Revised 12/12 32 26

18.5

M 1

Figure 1. SDS-PAGE analysis of Streptococcus faecalis NADH peroxidase expressed in E. coli. Electrophoresis was performed using a 10% polyacrylamide gel. Lane M, molecular weight marker; Lane 1, purified NADH peroxidase

(51 kDa).

Storage temperature

NADH peroxidase should be stored at 4 °C or and will remain stable up to 3 years if stored as specified. Certificate of Analysis

Test Criteria Result

Protein purity Purity in line with the stated value Meets specification

Protein concentration Concentration in line with the stated value Meets specification

Catalytic activity Activity in line with the stated value Meets specification

Blank assay variability Absorbance values with less than 10% of variability Meets specification

Approved by: José Prates Senior Manager, Quality Systems

Please enquire [email protected] to obtain any additional information. Bulk quantities of this product are available on request.

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