Microbial Metabolic Engineering: Methods and Protocols

ISBN-13: 9781493991419 / Angielski / Twarda / 2019 / 252 str.

Christine Santos; Ajikumar Parayil

Spis treści

1. Pathway and Gene Discovery from Natural Hosts and Organisms

Jing Ke and Yasuo Yoshikuni

 

2. Metabolic In silico Network Expansions to Predict and Exploit Enzyme Promiscuity

James Jeffryes, Jonathan Strutz, Chris Henry, and Keith E.J. Tyo

 

3. Antisense RNA Elements for Downregulating Expression

Yaping Yang, Jian Wang, Ruihua Zhang, and Yajun Yan

  4. Chemically Inducible Chromosomal Evolution (CIChE) for Multi-copy Metabolic Pathway Engineering

Aaron M. Love, Bradley W. Biggs, Keith E.J. Tyo, and Parayil Kumaran Ajikumar

 

5. Bacterial Expression of Membrane Associated Cytochrome P450s and their Activity Assay in Nanodiscs

Demetrios Maroutsos, Hannah Huff, and Aditi Das

 

6. Delta Integration CRISPR-Cas (Di-CRISPR) in Saccharomyces cerevisiae

Shuobo Shi, Youyun Liang, Ee Lui Ang, and Huimin Zhao

 

7. Assembly of Complex Pathways using Type IIs Restriction Enzymes

Sylvestre Marillonnet and Stefan Werner

  8. Combinatorial Assembly of Multigene Pathways by Combining Single Strand Assembly with Golden Gate Assembly D. Bauwens, P. Coussement, J. Maertens, and M. De Mey

 

9. Checks and Balances with Use of the Keio Collection for Phenotype Testing

Sandra J. Aedo, Helena R. Ma, and Mark P. Brynildsen

 

10. Genetic Engineering of Cyanobacteria: Design, Implementation, and Characterization of Recombinant Synechocystis sp. PCC 6803

Jacob Sebesta, Allison Zimont, and Christie Ann Marie Peebles

 

11. Genetic Tools for Streamlined and Accelerated Pathway Engineering in Yarrowia lipolytica

Lynn Wong, Benjamin Holdridge, Jake Engel, and Peng Xu

 

12. Gas Chromatography-mass Spectrometry Microbial Metabolomics for Applications in Strain Optimization

McKenzie L. Smith, April M. Miguez, and Mark P. Styczynski

 

13. Targeted Mass Spectrometry-based Proteomics Tools for Strain Optimization

Hsine-Chung Tseng and Christine Nicole S Santos

 

14. A Protocol for the Construction and Curation of Genome-scale Integrated Metabolic and Regulatory Network Models

Sriram Chandrasekaran

 

15. ¹³C-Fingerprinting and Metabolic Flux Analysis of Microbial Metabolisms  

Whitney Hollinshead, Lian He, and Yinjie J. Tang

 

16. A Genetic Screen for the Isolation of Mutants with Increased Flux in the Isoprenoid Pathway of Yeast

Manisha Wadhwa andAnand K Bachhawat

Blurb

This volume covers a wide array of topics that will aid researchers in the task of engineering complex biological systems. This book is divided into three parts: Part One discusses the discovery and identification of relevant biosynthetic pathways for engineering; Part Two looks at the development of genetic tools for manipulating enzymes, biosynthetic pathways, and whole genomes; and Part Three covers the characterization of engineered microbes using targeted and global systems biology tools, as well as in silico models. Chapters explore topics such as leveraging enzyme promiscuity to construct novel biosynthetic pathways; assembling combinatorial multigene pathways for rapid strain optimization; applying 'omics technologies for identifying bottlenecks; and engineering nontraditional host organisms like cyanobacterium and Yarrowia lipolytica. 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 authoritative, Microbial Metabolic Engineering: Methods and Protocols is a valuable resource for researchers and scientists interested in engineering and optimizing microbes for a variety of biotechnological applications.