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Flavins and Flavoproteins

Methods and Protocols

  • Book
  • © 2021

Overview

Editors:
  1. Maria Barile

    1. Department of Biosciences, Biotechnology and Biopharmaceutics, University of Bari “A. Moro”, Bari, Italy

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  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2280)

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Table of contents (18 protocols)

  1. Front Matter

    Pages i-xiii
    Download chapter PDF

  2. Riboflavin Production and Export by Microorganisms

    1. Front Matter

      Pages 1-1
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    2. Selection of Riboflavin Overproducing Strains of Lactic Acid Bacteria and Riboflavin Direct Quantification by Fluorescence

      • Pasquale Russo, Nicola De Simone, Vittorio Capozzi, Mari Luz Mohedano, José Ángel Ruiz-Masó, Gloria del Solar et al.
      Pages 3-14

    3. Recent Advances in Construction of the Efficient Producers of Riboflavin and Flavin Nucleotides (FMN, FAD) in the Yeast Candida famata

      • Dariya V. Fedorovych, Kostyantyn V. Dmytruk, Andriy A. Sibirny
      Pages 15-30

    4. Overexpression of Riboflavin Excretase Enhances Riboflavin Production in the Yeast Candida famata

      • Andriy O. Tsyrulnyk, Dariya V. Fedorovych, Kostyantyn V. Dmytruk, Andriy A. Sibirny
      Pages 31-42

  3. Measuring Riboflavin Transport and Flavin Cofactor Metabolism with Recombinant Human Proteins/Cell Fractions

    1. Front Matter

      Pages 43-43
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    2. Functional Study of the Human Riboflavin Transporter 2 Using Proteoliposomes System

      • Lara Console, Maria Tolomeo, Cesare Indiveri
      Pages 45-54

    3. Heterologous Overexpression of Human FAD Synthase Isoforms 1 and 2

      • Michele Galluccio, Cesare Indiveri
      Pages 55-67

    4. Purification of Recombinant Human 6His-FAD Synthase (Isoform 2) and Quantitation of FAD/Protein Monomer Ratio by UV-Vis Spectra

      • Piero Leone, Stefano Quarta, Maria Tolomeo, Maria Barile
      Pages 69-85

    5. Continuous and Discontinuous Approaches to Study FAD Synthesis and Degradation Catalyzed by Purified Recombinant FAD Synthase or Cellular Fractions

      • Piero Leone, Maria Tolomeo, Maria Barile
      Pages 87-116

  4. Looking at Flavin Cofactors in Flavoenzymes

    1. Front Matter

      Pages 117-117
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    2. Redox Titration of Flavoproteins: An Overview

      • Francesco Bonomi, Stefania Iametti
      Pages 119-133

    3. Anaerobic Stopped-Flow Spectrophotometry with Photodiode Array Detection in the Presteady State: An Application to Elucidate Oxidoreduction Mechanisms in Flavoproteins

      • Patricia Ferreira, Milagros Medina
      Pages 135-155

    4. Atomic Force Microscopy: Single-Molecule Imaging and Force Spectroscopy in the Study of Flavoproteins Ligand Binding and Reaction Mechanisms

      • Anabel Lostao, Milagros Medina
      Pages 157-178

    5. Ligand Binding in Allosteric Flavoproteins: Part 1. Quantitative Analysis of the Interaction with NAD+ of the Apoptosis Inducing Factor (AIF) Harboring FAD in the Reduced State

      • Paolo Cocomazzi, Luca Sorrentino, Federica Cossu, Alessandro Aliverti
      Pages 179-187

    6. Ligand Binding in Allosteric Flavoproteins: Part 2. Quantitative Analysis of the Redox-Dependent Interaction of the Apoptosis-Inducing Factor (AIF) with Its Protein Partner

      • Paolo Cocomazzi, Delia Tarantino, Eloise Mastrangelo, Alessandro Aliverti
      Pages 189-198

    7. Using d- and l-Amino Acid Oxidases to Generate the Imino Acid Substrate to Measure the Activity of the Novel Rid (Enamine/Imine Deaminase) Class of Enzymes

      • Stefania Digiovanni, Genny Degani, Laura Popolo, Maria Antonietta Vanoni
      Pages 199-218

    8. The In Vitro Production of prFMN for Reconstitution of UbiD Enzymes

      • Stephen A. Marshall, Karl Fisher, David Leys
      Pages 219-227

  5. Bioanalytical Applications of Flavoenzymes

    1. Front Matter

      Pages 229-229
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    2. Alcohol Oxidase from the Methylotrophic Yeast Ogataea polymorpha: Isolation, Purification, and Bioanalytical Application

      • Halyna M. Klepach, Andriy E. Zakalskiy, Oksana M. Zakalska, Galina Z. Gayda, Oleh V. Smutok, Mykhailo V. Gonchar
      Pages 231-248
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Keywords

About this book

This book of protocols is devoted to the yellow coenzymes derived from riboflavin or vitamin B2 and to the hundreds of enzymes whose functionality depends on them, and represents a compendium of techniques for working with flavoproteins or with the wide spectrum of functions that flavoproteins can drive in the cells. Starting with Rf production in microorganisms and the chemical, optical, and redox properties of these fascinating molecules and moving along to the variety and the peculiarity of some single flavoenzymes, the volume explores the complexity of functions and distribution of these molecules in the cell. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and practical, Flavin and Flavoproteins: Methods and Protocols serves as an ideal guide for protein chemists interested in purifying and characterizing flavoproteins, as well as microbiologists, physiologists, and clinicians, who wish to further study problems connected with flavoproteins.

Editors and Affiliations

  • Department of Biosciences, Biotechnology and Biopharmaceutics, University of Bari “A. Moro”, Bari, Italy

    Maria Barile

Bibliographic Information

  • Book Title: Flavins and Flavoproteins

  • Book Subtitle: Methods and Protocols

  • Editors: Maria Barile

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-1286-6

  • Publisher: Springer New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2021

  • Hardcover ISBN: 978-1-0716-1285-9Published: 23 March 2021

  • Softcover ISBN: 978-1-0716-1288-0Published: 23 March 2022

  • eBook ISBN: 978-1-0716-1286-6Published: 22 March 2021

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XIII, 298

  • Number of Illustrations: 28 b/w illustrations, 41 illustrations in colour

  • Topics: Enzymology, Chemistry/Food Science, general

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