Sarcosine Oxidase

PREPARATION and SPECIFICATION

PROPERTIES

APPLICATIONS

This enzyme is useful for enzymatic determination of creatinine when coupled with creatinine amidohydrolase and creatine amidohydrolase.

APPLICATIONS

UNIT DEFINITION

One unit (U) is defined as the amount of enzyme which produces 1 μmol of H2O2 per min under the conditions described below.

REAGENTS PREPARATION

Reagent I: 0.2 M Tris-HCl buffer, pH 8.0.

Reagent II: 1.0 M Sarcosine solution.

Reagent III: 1 kU/mL POD solution.

Reagent IV: 50 mM 4-AA solution.

Reagent V: 50 mM TOOS solution.

Enzyme diluent: 10 mM KH2PO4 – K2HPO4b uffer, pH 7.5.

Enzyme solution: Dilute the enzyme to 0.07 −0.17 U/ml with enzyme diluent.

REACTION MIXTURE (50 TESTS):

Reagent I                   5 mL

Reagent II                  10 mL

Reagent III                0.25 mL

Reagent IV                1.5 mL

Reagent V                  1.5 mL

Pure H2O                 31.75 mL

PROCEDURE

  1. Pipette 1 ml reaction mixture to a cuvette.
  2.  Preincubate the reaction mixture at 37℃ for 5 min.
  1.  Add 20 ul the enzyme solution and mix to start the reaction, record ΔAs at 555 nm in 1 minute in a spectrophotometer.

Measure the blank rate ΔAb by using the same method as the test except that the enzyme diluent is added instead of the enzyme solution.

∆A = ∆As – ∆Ab

CALCULATION

CALCULATION

REFERENCES

  1. N.Mori, M.Sato, Y.Tani and Y.Yamada; Agric.Biol.Chem., 44, 1391 (1980).
  1. M.Suzuki and M.Yoshida; Proceedings of the Symposium on Chemical Physiolosy and Pathology (Kyoto), Vol16, p.220 (1976).
  1. M.Suzuki; J. Biochem., 89, 599 (1981).