Pornpen SrisawasdiPatcharee JearanaikoonMartin H. KrollPorntip H. LolekhaMahidol UniversityKhon Kaen UniversityUT Southwestern Medical SchoolAsia Medical and Agricultural Laboratory and Research Center2018-06-212018-06-212005-12-12Journal of Clinical Laboratory Analysis. Vol.19, No.6 (2005), 247-252088780132-s2.0-28444449459https://repository.li.mahidol.ac.th/handle/20.500.14594/16257The enzymatic method for cholesterol determination can use either an endpoint or a kinetic method. Not much is known concerning the properties (Km and Vmax) of the commercial enzyme for the kinetic method. We measured the Km and Vmax of Brevibacterium, Streptomyces, Pseudomonas fluorescens, and Cellulomonas cholesterol oxidase. Brevibacterium gave the highest Km value (230.3 × 10 -4 M), followed by Streptomyces (2.17 × 10-4 M), Cellulomonas (0.84 × 10-4 M), and Pseudomonas (0.61 × 10-4 M). The Km values and the linearity obtained from Streptomyces (2.6 mmol/L), Pseudomonas (2.1 mmol/L), or Cellulomonas (2.1 mmol/L) were too low. Dichlorophenol isomers, acting as inhibitors, increased the enzyme's Km. The addition of 3,4-dichlorophenol raised the K m of Streptomyces from 2.17 × 10-4 to 24.89 × 10-4 M. The linearity was increased from 2.6 to 13.0 mmol/L. The high Km of Brevibacterium resulted in an insensitive reaction and low cholesterol linearity (7.8 mmol/L). An increase in the sample-to-reagent ratio from 1:100 to 1:10 enhanced the reaction rate and the linearity from 7.8 to 20.7 mmol/L. We suggest that Brevibacterium and Streptomyces cholesterol oxidase (with the addition of 3,4 dichlorophenol) are good sources for serum cholesterol determination by the kinetic method. © 2005 Wiley-Liss, Inc.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyHealth ProfessionsMedicineArticleSCOPUS10.1002/jcla.20086