Bookeditors: Vijai Singh,Ajay Kumar Singh,Poonam Bhargava, Madhvi Joshi, Chaitanya G. Joshi.
Summary: This book provides a comprehensive overview of the basic and advanced metabolic engineering technologies used to generate natural metabolites and industrially important biomolecules. Metabolic engineering has the potential to produce large quantities of valuable biomolecules in a renewable and sustainable manner by extending or modifying biosynthetic pathways in a wide range of organisms. It has been successfully used to produce chemicals, drugs, enzymes, amino acids, antibiotics, biofuels, and industrially important pharmaceuticals. The book comprehensively reviews the various metabolites detection, extraction and biosensors and the metabolic engineering of microbial strains for the production of industrially useful enzymes, proteins, organic acids, vitamins and antibiotics, therapeutics, chemicals, and biofuels. It also discusses various genetic engineering and synthetic biology tools for metabolic engineering. In closing, the book discusses ethical, patenting and regulatory issues in the metabolic engineering of microbes. This book is a valuable source not only for beginners in metabolic engineering, but also students, researchers, biotechnology and metabolic engineering based company.
Contents:
Intro
Foreword
Preface
Acknowledgments
Contents
About the Editors
1: An Introduction to Design of Microbial Strain using Synthetic Biology Toolboxes for Production of Biomolecules
1.1 Introduction
1.2 Desired Characteristics of Strain Used for Production of Biomolecules
1.3 Methods Used for Strain Improvement
1.4 Protein Engineering
1.5 Directed Evolution
1.6 Design and Construction of Biosynthetic Pathways
1.7 Synthetic Biology Toolbox in Metabolic Engineering
1.8 Adaptive Laboratory Evolution
1.9 Conclusion and Future Remarks
References 2.4.5 Engineering the Whole Cell
2.5 Genetic Engineering
2.5.1 Screening
2.5.2 Gene Expression
2.5.3 Enzyme (Over)Production and Posttranscriptional Control
2.6 Omics for Strain Engineering
2.6.1 Genome Analysis
2.6.2 Transcriptome Analysis
2.6.3 Proteome Analysis
2.6.4 Fluxome and Metabolome Analysis
2.6.4.1 Metabolome Analysis
2.6.4.2 Fluxome Analysis
2.6.5 Combined Omics Approach
2.7 Conclusion
References
3: Techniques for Detection and Extraction of Metabolites
3.1 Introduction
3.2 Extraction of Metabolites
3.2.1 Quenching Methods 2: Microbial Strain Engineering
2.1 Introduction
2.1.1 Need and Significance for Strain Improvement
2.2 Mutagenesis
2.2.1 Physical and Chemical Mutagenesis
2.2.2 Mutation Signature
2.3 Engineering Physiology of Microbes
2.3.1 Desired Physiological Characteristics
2.3.2 Candidate Screening
2.3.3 Selection of Host Strain
2.3.4 Engineering into the Host Strain
2.4 Metabolic Engineering
2.4.1 Methodologies and Tools for Metabolic Engineering
2.4.2 Engineering of Biosynthetic Pathways
2.4.3 Central Metabolism Engineering
2.4.4 Transport Engineering 3.2.1.1 Perchloric Acid
3.2.1.2 Liquid Nitrogen Method
3.2.1.3 Methanol Method
3.2.2 Extraction of Extracellular Metabolites
3.2.3 Extraction of Intracellular Metabolites
3.2.3.1 Non-mechanical Lysis
Boiling Ethanol
Cold Methanol
Buffered Methanol-Chloroform-Water
Hot Water
3.2.3.2 Mechanical Lysis of Cell
Supercritical Fluid Extraction
Pressurized Liquid Extraction
3.3 Detection of Metabolites
3.3.1 Mass-Spectrometry (MS)
3.3.2 Gas Chromatography-MS
3.3.3 Capillary Electrophoresis-MS (CE-MS)
3.3.4 Liquid Chromatography-MS (LC-MS) 3.3.5 Nuclear Magnetic Resonance
3.4 Conclusion
References
4: Genetically Encoded Biosensors and Their Applications in the Development of Microbial Cell Factories
4.1 The Classification of Genetically Encoded Biosensors
4.1.1 Protein-Based Biosensors
4.1.1.1 The Functional Mechanism of Protein-Based Biosensors
4.1.1.2 Designing and Tuning Protein-Based Biosensors
Tuning at aTF Level
Tuning at Promoter Level
4.1.2 RNA-Based Biosensors
4.1.2.1 The Functional Mechanism of RNA-Based Biosensors
4.1.2.2 Designing and Tuning RNA-Based Biosensors