Microbial Metabolic Engineering: Methods and Protocols

✍ Scribed by Christine Nicole S. Santos, Parayil Kumaran Ajikumar

• Publisher: Springer New York;Humana Press
• Year: 2019
• Tongue: English
• Leaves: 252
• Series: Methods in Molecular Biology 1927
• Edition: 1st ed.
• Category: Library

✦ Synopsis

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.

✦ Table of Contents

Front Matter ....Pages i-x
Pathway and Gene Discovery from Natural Hosts and Organisms (Jing Ke, Yasuo Yoshikuni)....Pages 1-9
Metabolic In Silico Network Expansions to Predict and Exploit Enzyme Promiscuity (James Jeffryes, Jonathan Strutz, Chris Henry, Keith E. J. Tyo)....Pages 11-21
Antisense RNA Elements for Downregulating Expression (Yaping Yang, Jian Wang, Ruihua Zhang, Yajun Yan)....Pages 23-35
Chemically Inducible Chromosomal Evolution (CIChE) for Multicopy Metabolic Pathway Engineering (Aaron M. Love, Bradley W. Biggs, Keith E. J. Tyo, Parayil Kumaran Ajikumar)....Pages 37-45
Bacterial Expression of Membrane-Associated Cytochrome P450s and Their Activity Assay in Nanodiscs (Demetrios Maroutsos, Hannah Huff, Aditi Das)....Pages 47-72
Delta Integration CRISPR-Cas (Di-CRISPR) in Saccharomyces cerevisiae (Shuobo Shi, Youyun Liang, Ee Lui Ang, Huimin Zhao)....Pages 73-91
Assembly of Complex Pathways Using Type IIs Restriction Enzymes (Sylvestre Marillonnet, Stefan Werner)....Pages 93-109
Combinatorial Assembly of Multigene Pathways by Combining Single-Strand Assembly with Golden Gate Assembly (D. Bauwens, P. Coussement, J. Maertens, Marjan De Mey)....Pages 111-123
Checks and Balances with Use of the Keio Collection for Phenotype Testing (Sandra J. Aedo, Helena R. Ma, Mark P. Brynildsen)....Pages 125-138
Genetic Engineering of Cyanobacteria: Design, Implementation, and Characterization of Recombinant Synechocystis sp. PCC 6803 (Jacob Sebesta, Allison Werner, Christie Ann Marie Peebles)....Pages 139-154
Genetic Tools for Streamlined and Accelerated Pathway Engineering in Yarrowia lipolytica (Lynn Wong, Benjamin Holdridge, Jake Engel, Peng Xu)....Pages 155-177
Gas Chromatography–Mass Spectrometry Microbial Metabolomics for Applications in Strain Optimization (McKenzie L. Smith, April M. Miguez, Mark P. Styczynski)....Pages 179-189
Targeted Mass Spectrometry-Based Proteomics Tools for Strain Optimization (Hsien-Chung Tseng, Christine Nicole S. Santos)....Pages 191-201
A Protocol for the Construction and Curation of Genome-Scale Integrated Metabolic and Regulatory Network Models (Sriram Chandrasekaran)....Pages 203-214
13C-Fingerprinting and Metabolic Flux Analysis of Bacterial Metabolisms (Whitney Hollinshead, Lian He, Yinjie J. Tang)....Pages 215-230
A Genetic Screen for the Isolation of Mutants with Increased Flux in the Isoprenoid Pathway of Yeast (Manisha Wadhwa, Anand K. Bachhawat)....Pages 231-246
Back Matter ....Pages 247-252

✦ Subjects

Biomedicine; Biomedical Engineering/Biotechnology; Microbiology