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  • © 2013

Enzyme Engineering

Methods and Protocols

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

Part of the book series: Methods in Molecular Biology (MIMB, volume 978)

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Table of contents (18 protocols)

  1. Front Matter

    Pages i-xi
  2. Determining Enzyme Kinetics via Isothermal Titration Calorimetry

    • Neil A. Demarse, Marie C. Killian, Lee D. Hansen, Colette F. Quinn
    Pages 21-30
  3. GFP Reporter Screens for the Engineering of Amino Acid Degrading Enzymes from Libraries Expressed in Bacteria

    • Olga Paley, Giulia Agnello, Jason Cantor, Tae Hyun Yoo, George Georgiou, Everett Stone
    Pages 31-44
  4. Flow Cytometric Assays for Interrogating LAGLIDADG Homing Endonuclease DNA-Binding and Cleavage Properties

    • Sarah K. Baxter, Abigail R. Lambert, Andrew M. Scharenberg, Jordan Jarjour
    Pages 45-61
  5. TAL Effector Nuclease (TALEN) Engineering

    • Ting Li, Bing Yang
    Pages 63-72
  6. In Vitro Evolution of Enzymes

    • Misha V. Golynskiy, John C. Haugner III, Aleardo Morelli, Dana Morrone, Burckhard Seelig
    Pages 73-92
  7. Residue-Specific Incorporation of Unnatural Amino Acids into Proteins In Vitro and In Vivo

    • Amrita Singh-Blom, Randall A. Hughes, Andrew D. Ellington
    Pages 93-114
  8. Reconstructing Evolutionary Adaptive Paths for Protein Engineering

    • Megan F. Cole, Vanessa E. Cox, Kelsey L. Gratton, Eric A. Gaucher
    Pages 115-125
  9. Gene Synthesis by Assembly of Deoxyuridine-Containing Oligonucleotides

    • Romualdas Vaisvila, Jurate Bitinaite
    Pages 165-171
  10. Protein Engineering: Single or Multiple Site-Directed Mutagenesis

    • Pei-Chung Hsieh, Romualdas Vaisvila
    Pages 173-186
  11. Gene Assembly and Combinatorial Libraries in S. cerevisiae via Reiterative Recombination

    • Nili Ostrov, Laura M. Wingler, Virginia W. Cornish
    Pages 187-203
  12. Promiscuity-Based Enzyme Selection for Rational Directed Evolution Experiments

    • Sandeep Chakraborty, Renu Minda, Lipika Salaye, Abhaya M. Dandekar, Swapan K. Bhattacharjee, Basuthkar J. Rao
    Pages 205-216
  13. In Vitro Directed Evolution of Enzymes Expressed by E. coli in Microtiter Plates

    • Bradley J. Stevenson, Sylvia H.-C. Yip, David L. Ollis
    Pages 237-249
  14. Back Matter

    Pages 251-252

About this book

Whether the pursuit is commercially motivated or purely academic, engineering a novel biological catalyst is an enticing challenge. High-resolution protein structure analysis allows for rational alteration of enzyme function, yet many useful enzyme variants are the product of well-designed selection schemes or screening strategies. Enzyme Engineering: Methods and Protocols provides guidance to investigators wishing to create enzyme variants with desired properties. This detailed volume covers such topics as a simple method for generating site-specific mutations within bacterial chromosomes. It also highlights the engineering of two difference types of rare-cutting endonucleases that show great potential in gene therapy applications: The newest development is the emergence of TAL effector nucleases or TALENs. Chapters describe newly developed technologies in sufficient detail so that each method can be practiced in a standard molecular biology laboratory. Written in the successful Methods in Molecular Biologyâ„¢ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

 

Authoritative and easily accessible Enzyme Engineering: Methods and Protocols will be valuable for scientists with a budding interest in protein engineering as well as veterans looking for new approaches to apply in established discovery programs.

Editors and Affiliations

  • New England Biolabs, Inc., Ipswich, USA

    James C. Samuelson

Bibliographic Information

  • Book Title: Enzyme Engineering

  • Book Subtitle: Methods and Protocols

  • Editors: James C. Samuelson

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-62703-293-3

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media New York 2013

  • Hardcover ISBN: 978-1-62703-292-6Published: 20 February 2013

  • Softcover ISBN: 978-1-4939-6287-7Published: 23 August 2016

  • eBook ISBN: 978-1-62703-293-3Published: 20 February 2013

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 252

  • Topics: Biochemistry, general, Enzymology

Buy it now

Buying options

eBook USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Other ways to access