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Protein Engineering

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Cover of 'Protein Engineering'

Table of Contents

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    Book Overview
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    Chapter 1 Protein Engineering: Past, Present, and Future
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    Chapter 2 Rational and Semirational Protein Design
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    Chapter 3 Computational Analysis of Protein Tunnels and Channels
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    Chapter 4 YASARA: A Tool to Obtain Structural Guidance in Biocatalytic Investigations
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    Chapter 5 A Computational Library Design Protocol for Rapid Improvement of Protein Stability: FRESCO
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    Chapter 6 Directed Evolution of Proteins Based on Mutational Scanning
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    Chapter 7 A Brief Guide to the High-Throughput Expression of Directed Evolution Libraries
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    Chapter 8 Library Growth and Protein Expression: Optimal and Reproducible Microtiter Plate Expression of Recombinant Enzymes in E. coli Using MTP Shakers
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    Chapter 9 Normalized Screening of Protein Engineering Libraries by Split-GFP Crude Cell Extract Quantification
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    Chapter 10 Functional Analysis of Membrane Proteins Produced by Cell-Free Translation
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    Chapter 11 Practical Considerations Regarding the Choice of the Best High-Throughput Assay
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    Chapter 12 High-Throughput Screening Assays for Lipolytic Enzymes
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    Chapter 13 Continuous High-Throughput Colorimetric Assays for α -Transaminases
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    Chapter 14 Colorimetric High-Throughput Screening Assays for the Directed Evolution of Fungal Laccases
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    Chapter 15 Directed Coevolution of Two Cellulosic Enzymes in Escherichia coli Based on Their Synergistic Reactions
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    Chapter 16 Program-Guided Design of High-Throughput Enzyme Screening Experiments and Automated Data Analysis/Evaluation
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    Chapter 17 Solid-Phase Agar Plate Assay for Screening Amine Transaminases
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    Chapter 18 Ultrahigh-Throughput Screening of Single-Cell Lysates for Directed Evolution and Functional Metagenomics
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    Chapter 19 Isolation of pH-Sensitive Antibody Fragments by Fluorescence-Activated Cell Sorting and Yeast Surface Display
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    Chapter 20 Library Generation and Auxotrophic Selection Assays in Escherichia coli and Thermus thermophilus
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    Chapter 21 Erratum to: Functional Analysis of Membrane Proteins Produced by Cell-Free Translation
Attention for Chapter 14: Colorimetric High-Throughput Screening Assays for the Directed Evolution of Fungal Laccases
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Chapter title
Colorimetric High-Throughput Screening Assays for the Directed Evolution of Fungal Laccases
Chapter number 14
Book title
Protein Engineering
Published in
Methods in molecular biology, January 2018
DOI 10.1007/978-1-4939-7366-8_14
Pubmed ID
Book ISBNs
978-1-4939-7364-4, 978-1-4939-7366-8
Authors

Isabel Pardo, Susana Camarero, Pardo, Isabel, Camarero, Susana

Abstract

In this chapter we describe several high-throughput screening assays for the evaluation of mutant libraries for the directed evolution of fungal laccases in the yeast Saccharomyces cerevisiae. The assays are based on the direct oxidation of three syringyl-type phenols derived from lignin (sinapic acid, acetosyringone, and syringaldehyde), an artificial laccase mediator (violuric acid), and three organic synthetic dyes (Methyl Orange, Evans Blue, and Remazol Brilliant Blue). While the assays with the natural phenols can be used for laccases with low redox potential, the rest are exclusive for high-redox potential laccases. In fact, the violuric acid assay is devised as a method to ascertain that the high-redox potential of laccase is not lost during directed evolution.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 14 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 14 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 4 29%
Student > Bachelor 2 14%
Other 1 7%
Lecturer 1 7%
Librarian 1 7%
Other 4 29%
Unknown 1 7%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 4 29%
Agricultural and Biological Sciences 3 21%
Unspecified 1 7%
Pharmacology, Toxicology and Pharmaceutical Science 1 7%
Environmental Science 1 7%
Other 2 14%
Unknown 2 14%