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Determination of Michaelis–Menten Enzyme Kinetics Parameters of Alkaline Phosphatase in Clinical Samples

  • Varsha GuptaEmail author
  • Abhishek Gupta
  • Lata I. Shukla
  • Abhimanyu Kumar Jha
  • Jaya Prakash
  • Baishnab Charan Tripathy
Protocol
  • 25 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Enzymes are specific biocatalysts which enhance the rate of biological reactions in mild conditions of temperature, pressure and a particular pH and thus have an important role in almost every cellular process. They play an indispensable role in metabolism, signal transduction cascade, and gene expression which makes them an important target for drug development, bioprocess industry, food industry, and eco-friendly green chemistries. They are emerging as potential drug targets. Many diseases are been explored where enzyme inhibition can have therapeutic effects. The study and analysis of enzyme have been challenging due to its mathematical aspects. However, due to their important functions, it becomes imperative to have sufficient and thorough knowledge of biochemistry of an enzyme and deep understanding of kinetics of enzyme action. In the present chapter, we will explore very simple and established basic concept of enzyme kinetics, estimation of initial reaction velocities with different substrate concentration, measurement and utility of KM and Vmax, catalytic power of the enzyme along with its turnover number. The readers will learn about changes which occur in their graph with different enzyme inhibitors in their reaction. This chapter would also be helpful for researchers and students to understand positive and negative feedback loop. We have provided example of serum alkaline phosphatase (ALP) where one can learn to establish different parameters with increase in absorbance as substrate concentration increases.

Keywords

Enzyme kinetics Michaelis–Menten equation Lineweaver–Burk plot Eadie–Hofstee plot Alkaline phosphatase Allosteric regulation 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Varsha Gupta
    • 1
    Email author
  • Abhishek Gupta
    • 2
  • Lata I. Shukla
    • 3
  • Abhimanyu Kumar Jha
    • 4
    • 5
  • Jaya Prakash
    • 6
  • Baishnab Charan Tripathy
    • 7
  1. 1.Rheumatology Laboratory, Department of BiotechnologyChhatrapati Shahu Ji Maharaj UniversityKanpurIndia
  2. 2.I3Consulting Pvt. Ltd. Sector 127NoidaIndia
  3. 3.Department of Biotechnology, School of Life SciencesPondicherry UniversityKalapetIndia
  4. 4.Institute of Technology and ManagementMeerutIndia
  5. 5.Department of Biotechnology, Faculty of Life SciencesInstitute of Applied Medicines and ResearchGhaziabadIndia
  6. 6.Community Health CentreKanpurIndia
  7. 7.School of Life SciencesJawaharlal Nehru UniversityNew DelhiIndia

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