With the advent of recombinant DNA technology, expressing heterologous proteins in microorganisms rapidly became the method of choice for their production at laboratory and industrial scale. Bacteria, yeasts and other hosts can be grown to high biomass levels efficiently and inexpensively. Obtaining high yields of recombinant proteins from this material was only feasible thanks to constant research on microbial genetics and physiology that led to novel strains, plasmids and cultivation strategies. Despite the spectacular expansion of the field, there is still much room for progress. Improving the levels of expression and the solubility of a recombinant protein can be quite challenging. Accumulation of the product in the cell can lead to stress responses which affect cell growth. Buildup of insoluble and biologically inactive aggregates (inclusion bodies) lowers the yield of production. This is particularly true for obtaining membrane proteins or high-molecular weight and multi-domain proteins. Also, obtaining eukaryotic proteins in a prokaryotic background (for example, plant or animal proteins in bacteria) results in a product that lack post-translational modifications, often required for functionality. Changing to a eukaryotic host (yeasts or filamentous fungi) may not be a proper solution since the pattern of sugar modifications is different than in higher eukaryotes. Still, many advances in the last couple of decades have provided to researchers a wide variety of strategies to maximize the production of their recombinant protein of choice. Everything starts with the careful selection of the host. Be it bacteria or yeast, a broad list of strains is available for overcoming codon use bias, incorrect disulfide bond formation, protein toxicity and lack of post-translational modifications. Also, a huge catalog of plasmids allows choosing for different fusion partners for improving solubility, protein secretion, chaperone co-expression, antibiotic resistance and promoter strength. Next, controlling culture conditions like temperature, inducer and media composition can bolster recombinant protein production. With this Research Topic, we aim to provide an encyclopedic account of the existing approaches to the expression of recombinant proteins in microorganisms, highlight recent discoveries and analyze the future prospects of this exciting and ever-growing field.

Recombinant Protein Expression, Part A, Volume 659 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more. Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Enzymology serial - Updated release includes the latest information on Recombinant Protein Expression

While the choices of microbial and eukaryotic expression systems for production of recombinant proteins are many, most researchers in academic and industrial settings do not have ready access to pertinent biological and technical information since it is normally scattered throughout the scientific literature. This book closes the gap by providing information on the general biology of the host organism, a description of the expression platform, a methodological section -- with strains, genetic elements, vectors and special methods, where applicable -- as well as examples of proteins produced with the respective platform. The systems thus described are well balanced by the inclusion of three prokaryotes (two Gram-negatives and one Gram-positive), four yeasts, two filamentous fungi and two higher eukaryotic cell systems -- mammalian and plant cells. Throughout, the book provides valuable practical and theoretical information on the criteria and schemes for selecting the appropriate expression platform, the possibility and practicality of a universal expression vector, and on comparative industrial-scale fermentation, with the production of a recombinant Hepatitis B vaccine chosen as an industrial example. With a foreword by Herbert P. Schweizer, Colorado State University, USA: "As a whole, this book is a valuable and overdue resource for a varied audience. It is a practical guide for academic and industrial researchers who are confronted with the design of the most suitable expression platform for their favorite protein for technical or pharmaceutical purposes. In addition, the book is also a valuable study resource for professors and students in the fields of applied biology and biotechnology."

The design and production of novel peptides and proteins occupy pivotal positions in science and technology and will continue to do so in the 21st century. Protein Engineering and Design outlines the rapid advances in computer-based modeling, protein engineering, and methods needed for protein and peptide preparation and characterization. This indispensable reference lays the groundwork for understanding this multidisciplinary activity while providing an introduction for researchers and students to the field of protein design. - Introduces and defines the techniques involved in protein engineering and design - Provides a concise overview of key technologies involved and demonstrates their contributions to the specialized design and production of novel proteins and peptides

Gram-negative Escherichia coli (E. coli) bacteria are the most numerous commensal aerobic germs located in the human colon. Diarrhea caused by E. coli pathogenic strains is a major cause of death in developing countries, especially the sub-Saharan and South Asian areas. Some strains cause diarrhea, and all of them may produce an infectious disease. This book includes ten chapters covering the main aspects of infections related to E. coli, their pathogenic mechanisms, treatments, and resistance to diverse antibiotics.

New and Future Developments in Microbial Biotechnology and Bioengineering: Recent Advances in Application of Fungi and Fungal Metabolites: Biotechnological Interventions and Futuristic Approaches is an invaluable resource for researchers planning to work in applied biotechnological interventions and futuristic approaches to fungi and fungal metabolite utilization. Special emphasis is placed on new research relating to fungal-based recombinant DNA technology and genomics analysis which place yeasts and filamentous fungi at the forefront of various contemporary commercial applications. Written in an easy-to-follow language by active researchers, the book presents cutting-edge fungal biotechnological applications in a manner that is accessible to all. Introduces recent biotechnological interventions and futuristic approaches to fungi and their metabolites Elaborates on perspectives and diverse applications of harnessing the potential of fungi and fungal metabolites in biotechnology Describes traditional uses and modern practices of accessing the potential of fungi and their metabolites in solving future needs

This two-volume set discusses recent approaches and technological innovations for sustainable agriculture in smallholder farming systems impacted by climate change. The systems covered include crop-based agricultural production, as well as aquaculture and livestock production as related systems using similar techniques to combat food security issues brought about by climate change and resource overuse. The chapters detail innovations involving crop diversification, soil resilience management, geoinformatics and land suitability monitoring for smart farming, information technology in livestock production, and nutrient resource management in fishery aquaculture. Researchers, practitioners and industries will be able to use this information to implement socially and economically sustainable practices to achieve food security in impoverished areas vulnerable to climate change, while also learning about the rapid evolution in information technology that is applicable for and available to small holder farmers. Volume 2 focuses on trends and technologies in food security within the context of sustainable practices, drone technology, microwave data, molecular farming, machine learning, agricultural economics, spatial modeling and agricultural policy. These chapters discuss advancements in fishery resource and aquaculture practices, and also the challenges facing these areas due to climate change.

Biotechnology has emerged as one of the key environmentally safe technologies for the future which enables use of biomass to develop novel smart materials and to replace oil derived products. Fungi are the most efficient producers of the enzymes needed for this purpose and in addition they produce a plethora of secondary metabolites, among which novel antibiotics can be found. Industrial application and exploitation of the metabolic capacities of fungi requires highly productive and robust gene expression systems, which can be achieved by selection of appropriate species and strain improvement. In this book we aim to summarize homologous and heterologous gene expression systems of fungi for production of enzymes and secondary metabolites. A broad overview on requirements, challenges and successful applications shall serve as a basis for further development of fungi as biotechnological workhorses in research and industry.

This edited volume emphasizes how microorganisms have become a reliable pillar of biotechnology. The authors discuss advances in synthetic biology and genetic engineering that have made it possible to reprogram the microbial cellular capabilities. This enables an economically viable production of high-value products at an industrial level. The first part of the book provides an overview of synthetic biology and genome editing tools for engineering microbial cell factories in modern fermentation. Readers also learn how high-throughput bioprocessing methods are used to recover and purify microbial products. The remaining parts of this book explore the implementation and challenges of these upstream and downstream processing techniques for manufacturing high-value products. Cost-effectiveness and quality-control are key factors, when discussing the production of low-molecular-weight products, biopharmaceuticals, biopolymers and protein-based nanoparticles. This book is a valuable resource for biotechnologists both in the industry and in academia.