Development and Evaluation of Kinetic Spectrophotometric Assays for Horseradish Peroxidase by Catalytic Coupling of Paraphenylenediamine and Mequinol

• NAGARAJA Padmarajaiah

  Department of Studies in Chemistry, University of Mysore

• SHIVAKUMAR Anantharaman

  Department of Studies in Chemistry, University of Mysore

• KUMAR SHRESTHA Ashwinee

  Department of Studies in Chemistry, University of Mysore

抄録

This paper presents a novel spectrophotometric method to measure peroxidase activity using paraphenylenediamine dihydrochloride (PPDD) and Mequinol (MQ). The PPDD traps the free radical, and gets oxidized to electrophilic 1,4-diimine; this couples with MQ to an give intense violet-colored chromogenic species with the maximum absorbance at 560 nm. This assay was adopted for the quantification of hydrogen peroxide between 10 × 10⁻⁶ to 80 × 10⁻⁶ M. From the kinetic data, a two-substrate ping-pong mechanism of peroxidase was established. Catalytic efficiency and catalytic power of commercial peroxidase were 0.204 × 10⁶ M⁻¹ min⁻¹ and 2.86 × 10⁻⁴ min⁻¹, respectively. The catalytic constant (k_cat) of the proposed method was 0.2080 × 10³ min⁻¹. As a simple, rapid, precise and sensitive technique, PPDD-MQ stands as a potential replacement for the traditional guaiacol method. Applications to the plant extracts increase its relevance in the field of biochemical analysis.

収録刊行物

• Analytical Sciences

  Analytical Sciences 25 (10), 1243-1248, 2009

  社団法人 日本分析化学会

キーワード

• Analytical Chemistry

詳細情報

• CRID: 1390282679234145792
• NII論文ID: 130004441768
• NII論文ID: 10025623346
• NII書誌ID: AA10500785
• DOI: 10.2116/analsci.25.1243
• ISSN: 13482246
• ISSN: 09106340
• NDL書誌ID: 10384807
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I10384807
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.25.1243.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.25.1243/fulltext.html
• Web Site: http://www.jstage.jst.go.jp/article/analsci/25/10/25_10_1243/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可