Publication: Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol
Issued Date
2005-12-12
Resource Type
ISSN
08878013
Other identifier(s)
2-s2.0-28444449459
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Journal of Clinical Laboratory Analysis. Vol.19, No.6 (2005), 247-252
Suggested Citation
Pornpen Srisawasdi, Patcharee Jearanaikoon, Martin H. Kroll, Porntip H. Lolekha Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol. Journal of Clinical Laboratory Analysis. Vol.19, No.6 (2005), 247-252. doi:10.1002/jcla.20086 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/16257
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol
Abstract
The 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.