Main description:

This volume discusses the latest research and techniques used to study novel cellulases. The chapters in this book explore topics such as the role of cellulases in lignocellulose deconstruction for generation of a sugar platform for biofuel; screening for novel fungal b-glucosidases in a variety of fungal strains; discovery of novel enzymes in bacteria or fungi; protocols for isolating cellulolytic rumen bacteria; analysis of enzyme mixtures produced by ligocellulolytic fungi; and homology modeling. Written in the highly successful Methods in Molecular Biology series format, 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.

Cutting-edge and thorough, Cellulases: Methods and Protocols is a valuable resource for biochemists, molecular biologists, chemical engineers, and industrial researchers interested in this field.

Contents:

Preface...

Table of Contents...

Contributing Authors...

Part I Introduction Chapter and a Case Example

1. Cellulases: Role in Lignocellulosic Biomass Utilization

Sanjeev Kumar Soni, Amita Sharma, and Raman Soni

2. Discovery of a Novel Fungus with an Extraordinary b-Glucosidase

Peter Stephensen Lubeck and Mette Lubeck

Part II Protocols

Discovery of New Cellulases Using Classical or Molecular Methods

3. Isolation and Screening of Cellulolytic Filamentous Fungi

Mette Lubeck and Peter Stephensen Lubeck

4. Isolation and Screening of Cellulose Degrading Microorganisms from Different Ecological Niches

Ayyappa Kumar Sista Kameshwar and Wensheng Qin

5. Isolation of Cellulolytic Bacteria from the Rumen

Makoto Mitsumori

6. Methods for Discovery of Novel Cellulosomal Cellulases using Genomics and Biochemical Tools

Yonit Ben-David, Bareket Dassa, Lizi Bensoussan, Edward A. Bayer, and Sarah Morais

7. Metatranscriptomic Techniques for Identifying Cellulases in Termites and Their Symbionts

Brittany F. Peterson and Michael E. Scharf

8. Discovery of Novel Cellulases Using Proteomic Strategies

Marta Zoglowek, Heather Brewer, and Angela Norbeck

9. Identification of Key Components for the Optimization of Cellulase Mixtures Using a Proteomic Strategy

Jingyao Qu, Jing Zhu, Guodong Liu, and Yinbo Qu

10. Cloning and Expression of Heterologous Cellulases and Enzymes in Aspergillus niger

Morgann C. Reilly, Saori Amaike Campen, Scott E. Baker, John M. Gladden, and Jon K. Magnuson

11. Advanced Cloning Tools for Construction of Designer Cellulosomes

Amaranta Kahn, Edward A. Bayer, and Sarah Morais

12. Evaluation of Thermal Stability of Cellulosomal Hydrolases and Their complex Formation

Amaranta Kahn, Anastasia P. Galanopoulou, Dimitris G. Hatzinikolaou, Sarah Morais, and Edward A. Bayer

Assays for Activity Testing

13. Endoglucanase (EG) Activity Assays

M. Shafiqur Rahman, Sheran Fernando, Brian Ross, Jiangning Wu, and Wenshing Qin

14. Cellobiohydolase (CBH) Activity Assays

Hem Kanta Sharma, Wensheng Qin, and Chunbao (Charles) Xu

15. Two-Dimensional High-Throughput Endo-Enzyme Screening Assays Based on Chromogenic Polysaccharide Hydrogel and Complex Biomass Substrates

Julia Schuckel and Stjepan Kresimir Kracun

16. Analytical Tools for Characterizing Cellulose-Active Lytic Polysaccharide Monooxygenases (LPMOs)

Bjorge Westereng, Jennifer S.M. Loose, Gustav Vaaje-Kolstad, Finn L. Aachmann, Morten Sorlie, and Vincent G.H. Eijsink

17. Light Induced Electron Transfer Protocol for Enzymatic Oxidation of Polysaccharide

David Cannella

18. Purification of Cellulase from Cellulolytic Microorganisms and Characterization of the Total Cellulase Activities (TCA)

Ayyappa Kumar Sista Kameshwar and Wensheng Qin

Application of Cellulases for Lignocellulose Degradation

19. Fungal On-Site Production of Cellulolytic Enzymes by the Sequential Cultivation Method

Cristiane S. Farinas, Camila Florencio, and Alberto C. Badino

20. Test of Efficacy of Cellulases for Biomass Degradation

Henning Joergensen

Bioinformatics for Improving Cellulases

21. Homology Modeling for Enzyme Design

Wimal Ubhayasekera