Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol
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 Cell...
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th-mahidol.162572018-06-21T15:18:45Z Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol 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 Biochemistry, Genetics and Molecular Biology Health Professions Medicine 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. 2018-06-21T08:07:32Z 2018-06-21T08:07:32Z 2005-12-12 Article Journal of Clinical Laboratory Analysis. Vol.19, No.6 (2005), 247-252 10.1002/jcla.20086 08878013 2-s2.0-28444449459 https://repository.li.mahidol.ac.th/handle/123456789/16257 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=28444449459&origin=inward |
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Biochemistry, Genetics and Molecular Biology Health Professions Medicine Pornpen Srisawasdi Patcharee Jearanaikoon Martin H. Kroll Porntip H. Lolekha Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol |
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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. |
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Mahidol University |
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Mahidol University Pornpen Srisawasdi Patcharee Jearanaikoon Martin H. Kroll Porntip H. Lolekha |
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Article |
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Pornpen Srisawasdi Patcharee Jearanaikoon Martin H. Kroll Porntip H. Lolekha |
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Pornpen Srisawasdi |
title |
Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol |
title_short |
Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol |
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Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol |
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Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol |
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Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol |
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performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol |
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2018 |
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https://repository.li.mahidol.ac.th/handle/123456789/16257 |
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