Magnani Francesca, Marabelli Chiara, Paradisi Francesca (eds.) Enzyme Engineering: Methods and Protocols

Humana Press, 2022. — 358 p. — (Methods in Molecular Biology 2397). — ISBN 978-1-0716-1825-7.This book provides comprehensive methods and protocols about enzyme design. The chapters are grouped by main topic, starting with methodologies describing library preparation and screening, state of the art techniques in directed evolution and rational design, followed by examples of immobilization of enzymes on sustainable polymers, as well as biocatalytic conversions mediated by homogenous enzymatic preparations or whole cells. 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 timely, Enzyme Engineering: Methods and Protocols is an ideal guide for both the novice and the veteran researcher interested in biocatalysis.Enzyme Libraries Preparation and Screening
Preparation of Soil Metagenome Libraries and Screening for Gene-Specific Amplicons
Ultrahigh-Throughput Screening of Metagenomic Libraries Using Droplet Microfluidics
Synthetic DNA Libraries for Protein Engineering Toward Process Improvement in Drug Synthesis
Directed Evolution
In Silico Prediction Methods for Site-Saturation Mutagenesis
Recombination of Compatible Substitutions by 2GenReP and InSiReP
Semi-rational and De Novo Design
Using the Evolutionary History of Proteins to Engineer Insertion-Deletion Mutants from Robust, Ancestral Templates Using Graphical Representation of Ancestral Sequence Predictions (GRASP)
Resurrecting Enzymes by Ancestral Sequence Reconstruction
Expression and In Vivo Loading of De Novo Proteins with Tetrapyrrole Cofactors
Rational Design
Rational-Design Engineering to Improve Enzyme Thermostability
Using Molecular Simulation to Guide Protein Engineering for Biocatalysis in Organic Solvents
In Silico Engineering of Enzyme Access Tunnels
Quantum-Mechanical/Molecular-Mechanical (QM/MM) Simulations for Understanding Enzyme Dynamics
Computational Enzyme Design at Zymvol
Biocatalytic Process Development
Advanced Enzyme Immobilization Technologies: An Eco-friendly Support, a Polymer-Stabilizing Immobilization Strategy, and an Improved Cofactor Co-immobilization Technique
Chemical Reaction Engineering to Understand Applied Kinetics in Free Enzyme Homogeneous Reactors
Enzymatic Oxidative Cascade for Oxofunctionalization of Fatty Acids in One-Pot
CRISPR-Cas9 Editing of the Synthesis of Biodegradable Polyesters Polyhydroxyalkanaotes (PHA) in Pseudomonas putida KT2440