Juan M Bolivar (editor) - Immobilization of Enzymes and Cells : Methods and Protocols

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For over 35 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimedMethods in Molecular Biologyseries. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice. These hallmark features were introduced by series editor Dr. John Walker and constitute the key ingredient in each and every volume of theMethods in Molecular Biologyseries. Tested and trusted, comprehensive and reliable, all protocols from the series are indexed in PubMed.

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The global industrial enzyme market is expected to grow from 5.5 billion in 2018 to $7.0 billion in 2023, according to Business Communication Company Research (BCC). This aligns with the increasing trend in the industry to develop more sustainable and economic processes. The use of enzymes in chemistry underpins the transition of our society toward a more ecological economy since enzymes catalyze chemical reactions in a very sustainable and efficient manner.

Unfortunately, the features underlying the biological origins of enzymes hamper their exploitation as industrial catalysts. Hence, scientists have been encouraged to develop new tools to improve enzymes for their industrial use. Enzyme immobilization is a necessary step to design enzyme reactors; however, the use of these necessary immobilization techniques to improve enzyme properties (e.g., stability) is the main interest nowadays. In this way, soluble and instable enzyme catalysts may be converted into highly stable and heterogeneous biocatalysts that are useful in improving reaction and reactor engineering. The fourth edition of this volume is focused on the aforementioned areas.

The science of enzyme immobilization demands new protocols to fabricate heterogeneous biocatalysts with better properties than the soluble enzymes under reaction conditions with industrial relevance. This important task requires combining knowledge from material sciences, surface chemistry, protein chemistry, biophysics, molecular biology, biocatalysis, and chemical engineering.

The fourth edition ofImmobilization of Enzymes and Cellsaims to complement and update previous editions as a useful guide to the design of biocatalysts. This book collects a diversity of contributions from distinguished professors and researchers that cover several topics in the field of enzyme and cell immobilization.

The book begins with an overview chapter discussing the most relevant aspects of the science of enzyme immobilization. Briefly, we emphasize those immobilization techniques that promote the improvement of enzyme properties. For example, multivalent attachments between enzyme and carriers surface to yield highly stable heterogeneous biocatalysts. In this overview, we stress the importance of protein orientation and the valency of the attachment to achieve a structural rigidification of the immobilized enzymes. Chaptersestablishes some guidelines on the main parameters necessary to characterize immobilized enzymes and their performances under the operational conditions.

Detailed protocols of different functionalization chemistries for the modulation of the immobilized enzyme properties (e.g., by controlling enzyme orientation and degree of multipoint attachment) are presented in Chapters, respectively.

Beyond standard methods and solid materials, this book describes protocols using new immobilization methodologies and emerging materials (Chapters, respectively.

Beyond the techniques and the materials, Chapteraddresses a specific aspect related to the co-immobilization of multienzyme systems, paying particular attention to the effect of protein colocalization on the performance of the biocatalytic system. This chapter identifies the cofactor-dependent multienzyme systems as ideal system to control enzyme spatial organization where recycling efficiency is crucial.

In the front line of this book, forecasting new trends in enzyme and cell immobilization, Chaptersdescribes an innovative methodology for rapid preparation of heterogeneous biocatalysts. This technology combines one-pot cell-free protein synthesis and immobilization in a single test tube for the manufacture of biomaterials, immobilized enzymes among them.

This book also highlights the development of new analytical techniques for the characterization of heterogeneous biocatalysts to have a better understanding of the science of immobilization. Chapterdescribes the use of solid-state NMR to understand enzyme immobilization at the molecular level through identifying conformational changes.