Microbial Metabolic Engineering : Methods and Protocols

Table of Contents

Preface

Preamble

1 Screening for Cellulases with Industrial Value and their use in Biomass Conversion

Julia Jürgensen, Nele Ilmberger and Wolfgang R. Streit

2 Reversal of NAD(P)H Cofactor Dependence by Protein Engineering

Sabine Bastian and Francis H. Arnold

3 Quantifying Plasmid Copy Number to Investigate Plasmid Dosage Effects Associated with Directed Protein Evolution

Samuel Million-Weaver, David L. Alexander, Jennifer M. Allen, and Manel Camps

4 High Isoprenoid Flux Escherichia coli as a Host for Carotenoids Production

Wonchul Suh

5 Mutagenic Inverted Repeats Assisted Genome Engineering (MIRAGE) in Saccharomyces cerevisiae: Deletion of gal7

Nikhil U. Nair and Huimin Zhao

6 Creation of New Metabolic Pathways or Improvement of Existing Metabolic Enzymes by in vivo Evolution in Escherichia coli

Isabelle Meynial-Salles and Philippe Soucaille

7 Bioluminescent Reporter Genes for Promoter Discovery

Tina K. Van Dyk

8 Recombination-based DNA Assembly and Mutagenesis Methods for Metabolic Engineering

Xiquan Liang, Lansha Peng, Billyana Tsvetanova, Ke Li, Jian-Ping Yang, Tony Ho, Josh Shirley, Liewei Xu, Jason Potter, Wieslaw Kudlicki, Todd Peterson, and Federico Katzen

9 Ethanol-tolerant Gene Identification in Clostridium thermocellum using Pyro-resequencing for Metabolic Engineering

Shihui Yang, Dawn M. Klingeman, and Steven D. Brown

10 Use of Proteomic Tools in Microbial Engineering for Biofuel Production

Shaomin Mao, Kaizhi Jia, Yanping Zhang, Yin Li

11 Metabolic Engineering of Antibiotic-producing Actinomycetes using in vitro Transposon Mutagenesis

Andrew R. Reeves and J. Mark Weber

12 Use FACS Sorting in Metabolic Engineering of Escherichia coli for Increased Peptide Production

Qiong Cheng, Kristin Ruebling-Jass, Jianzhong Zhang, Qi Chen and Kevin Croker

13 Using Flux Balance Analysis to Guide Microbial Metabolic Engineering

Kathleen A. Moran, Nathan C. Crook and Hal S. Alper

14 Using an Advanced Microfermentor System for Strain Screening and Fermentation Optimization

Dongming Xie

15 Rapid Strain Evaluation using Dynamic DO-stat fed-batch Fermentation Under Scale-down Conditions

Jun Sun

16 Preparation and Evaluation of Lignocellulosic Biomass Hydrolysates for Growth by Ethanolgenic Yeasts

Ying Zha, Ronald Slomp, Johan van Groenestijn and Peter Punt

17 Engineering Whole-cell Biosensors with No Antibiotic Markers for Monitoring Aromatic Compounds in the Environment

Aitor de las Heras and Víctor de Lorenzo

18 Metabolic Engineering for Acetate Control in Large Scale Fermentation

Yong Tao, Qiong Cheng and Alexander D, Kopatsis

19 Minimization and Prevention of Phage Infections in Bioprocesses

Marcin Los

Shaomin Mao, Kaizhi Jia, Yanping Zhang, Yin Li

11 Metabolic Engineering of Antibiotic-producing Actinomycetes using in vitro Transposon Mutagenesis

Andrew R. Reeves and J. Mark Weber

12 Use FACS Sorting in Metabolic Engineering of Escherichia coli for Increased Peptide Production

Qiong Cheng, Kristin Ruebling-Jass, Jianzhong Zhang, Qi Chen and Kevin Croker

13 Using Flux Balance Analysis to Guide Microbial Metabolic Engineering

Kathleen A. Moran, Nathan C. Crook and Hal S. Alper

14 Using an Advanced

Metabolic engineering is the practice of genetically optimizing metabolic and regulatory networks within cells to increase production and/or recovery of certain substance from cells. In Microbial Metabolic Engineering: Methods and Protocols expert researchers in the field detail many of the methods which are now commonly used to study metabolic engineering. These include methods and techniques to engineer genes and pathways, use of modern biotechnology tools in microbial metabolic engineering, and examples of metabolic engineering for real world applications such as whole cell biosensors and acetate control in large scale fermentation. 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 key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Microbial Metabolic Engineering: Methods and Protocols seeks to provide researchers with an overview of key topics on microbial metabolic engineering.