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Contents

List of Contributors XIII
Preface XXI

1 Directed Evolution of Ligninolytic Oxidoreductases: from Functional
Expression to Stabilization and Beyond 1
Eva Garcia-Ruiz, Diana M. Mate, David Gonzalez-Perez, Patricia
Molina-Espeja, Susana Camarero, Angel T. Mart´ınez, Antonio O.
Ballesteros, and Miguel Alcalde
1.1 Introduction 1
1.2 Directed Molecular Evolution 1
1.3 The Ligninolytic Enzymatic Consortium 3
1.4 Directed Evolution of Laccases 6
1.4.1 Directed Evolution of Low-Redox Potential Laccases 7
1.4.2 Directed Evolution of Medium-Redox Potential Laccases 7
1.4.3 Directed Evolution of Ligninolytic High-Redox Potential Laccases
(HRPLs) 8
1.5 Directed Evolution of Peroxidases and Peroxygenases 11
1.6 Saccharomyces cerevisiae Biomolecular Tool Box 15
1.7 Conclusions and Outlook 16
Acknowledgments 17
Abbreviations 17
References 18

2 New Trends in the In Situ Enzymatic Recycling of NAD(P)(H)
Cofactors 23
Erica Elisa Ferrandi, Daniela Monti, and Sergio Riva
2.1 Introduction 23
2.2 Recent Advancements in the Enzymatic Methods for the Recycling of
NAD(P)(H) Coenzymes and Novel Regeneration Systems 24
2.2.1 In Situ Regeneration of Reduced NAD(P)H Cofactors 24
2.2.1.1 Formate Dehydrogenase and Glucose Dehydrogenase 24
2.2.1.2 Phosphite Dehydrogenase 26
2.2.1.3 Hydrogenase 27