Stabilized enzymatic reagents for measuring glucose, creatine kinase and gamma-glutamyltransferase with thermostable enzymes from a thermophile, Bacillus stearothermophilus.
Stabilized enzymatic reagents for measuring some components in biological fluids have been successfully developed based on two kinds of thermostable enzymes derived from Bacillus stearothermophilus with separation of the reagent into two complementary solutions. The thermostable glucokinase produced was applied to the measurement of glucose and creatine kinase activity, while the alanine dehydrogenase produced was used for the measurement of gamma-glutamyltransferase activity. The enzymatic reagents were also stabilized by developing two separate reagents with an optimum pH for the main reagent components. The stability of the reagents in liquid form was examined at 10 degrees C. It was clearly shown that the reagents for measuring glucose and creatine kinase activity were stable and retained their full capability for accurate measurement in biological fluids for over one year. The alanine dehydrogenase product was stable for at least 40 days.
Tomita
K
K
College of Engineering, Kanto Gakuin University, Yokohama, Japan.
Nomura
K
K
Kondo
H
H
Nagata
K
K
Tsubota
H
H
D050788:Alanine Dehydrogenase; D000594:Amino Acid Oxidoreductases / Q000737:chemistry; D003402:Creatine Kinase / Q000032:analysis*; D004355:Drug Stability; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D005941:Glucokinase / Q000737:chemistry; D005947:Glucose / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D013696:Temperature; D005723:gamma-Glutamyltransferase / Q000032:analysis*
D007202:Indicators and Reagents; D000594:Amino Acid Oxidoreductases; D050788:Alanine Dehydrogenase; D005723:gamma-Glutamyltransferase; D005941:Glucokinase; D003402:Creatine Kinase; D005947:Glucose