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Please use this identifier to cite or link to this item: http://repository.li.mahidol.ac.th/dspace/handle/123456789/16257
Title: Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol
Authors: Pornpen Srisawasdi
Patcharee Jearanaikoon
Martin H. Kroll
Porntip H. Lolekha
Mahidol University
Khon Kaen University
UT Southwestern Medical School
Asia Medical and Agricultural Laboratory and Research Center
Keywords: Biochemistry, Genetics and Molecular Biology;Health Professions;Medicine
Issue Date: 12-Dec-2005
Citation: Journal of Clinical Laboratory Analysis. Vol.19, No.6 (2005), 247-252
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.
URI: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=28444449459&origin=inward
http://repository.li.mahidol.ac.th/dspace/handle/123456789/16257
ISSN: 08878013
Appears in Collections:Scopus 2001-2005

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