Cell-Free Production : System Development

This book reviews the development of cell-free production platforms and offers an authoritative perspective of the latest advances and methodologies in cell-free production systems. Readers will discover the biomanufacturing potential of in vitro biotransformation (ivBT) employing purified cascade multi-enzymes, the development of hydrogel-based multi-enzymatic systems for biosynthesis, and novel insights into the optimization of biocatalytic processes. Additionally, the book explores the cell-free production and regeneration of cofactors, shedding light on strategies to enhance the efficiency and sustainability of cellular processes.

In this book, particular attention is given to the progress of cell-free in vitro evolution techniques for optimizing enzyme performance, and the book also presents the integration of rapid and finely-tuned expression systems for deployable sensing applications, revolutionizing the field of biosensing. The synthesis and electrophysiological analysis of multipass voltage-gated ion channels tethered in microsomal membranes are explored, providing a deep understanding of cellular function at the molecular level. Lastly, the book covers compartmentalized cell-free expression systems for building synthetic cells, showcasing the potential for constructing artificial cellular systems with unique functionalities.

Given its breadth, this book appeals to academics, researchers, and professionals interested in the forefront of biotechnology, and together with the companion volume “Cell-free Macromolecular Synthesis”, both books highlight the research progresses on the basic and applied research of cell-free production systems in the last few years, being invaluable resources in the field.

Chapter “Cell-free synthesis and electrophysiological analysis of multipass voltage-gated ion channels tethered in microsomal membranes” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Yuan Lu is an Associate Professor at the Department of Chemical Engineering, Tsinghua University, China. After receiving B.S. (2004) and Ph.D. (2009) in chemical engineering from Tsinghua University, China, Dr Lu received postdoctoral training from Johns Hopkins University (2009-2010) and Stanford University (2010-2014), USA. He was a project researcher from 2014 to 2016 at the University of Tokyo, Japan. The central paradigm of Dr Lu’s research is using interdisciplinary approaches in engineering, biology, chemistry, physics, and computer science to manipulate biomolecules and complex biological systems. The main focus lies on developing and applying synthetic biology and other cutting-edge technologies to engineer nucleic acids, proteins, pathways, and networks for addressing most daunting challenges in human health.

Michael Jewett is the Walter P. Murphy Professor of Chemical and Biological Engineering and Director of the Center for Synthetic Biology at Northwestern University, USA. Dr Jewett received his PhD in 2005 from Stanford University, USA, completed postdoctoral studies at the Center for Microbial Biotechnology in Denmark and the Harvard University Medical School, and was a guest professor at the Swiss Federal Institute of Technology (ETH Zurich). He is the recipient of the NIH Pathway to Independence Award, David and Lucille Packard Fellowship in Science and Engineering, Camille-Dreyfus Teacher-Scholar Award, and a Finalist for the Blavatnik National Awards for Young Scientists, among others. He is the co-founder of SwiftScale Biologics, Stemloop Inc., Pearl Bio, Induro Therapeutics, and Design Pharmaceuticals. Jewett is a Fellow of AIMBE, AAAS, and NAI. Rather than attempt to balance the tug-of-war that exists between the cell’s physiological and evolutionary objectives on one side and the engineer’s process objectives on the other, Dr Jewett’s group is developing new strategies that widen the aperture of the traditional model of biotechnology. In one direction, the group seeks to create a new paradigm for engineering biocatalytic systems using cell-free biology. The foundational principle is that one can conduct precise, complex biomolecular transformations in crude lysates without using intact cells. This concept represents a significant departure from cell-based processes that rely on microscopic cellular ‘reactors,’ and provides an unprecedented and otherwise unattainable freedom of design to modify and control biological systems. Dr Jeweet’s groups is also pioneering new directions to repurpose the translation apparatus for synthetic biology. The goal is to monitor, interrogate, and understand the process of translation, and with this knowledge diversify, evolve and repurpose the ribosome and its peripheral machinery into a re-engineered machine to generate non-natural polymers as new classes of sequence-defined, evolvable matter.