Cell-Free Biosystems for Biomanufacturing
Although cell-free biosystems have been used as a tool for investigating fundamental aspects of biological systems for more than 100 years, they are becoming an emerging biomanufacturing platform in the production of low-value biocommodities (e.g., H2, et
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Cell-Free Biosystems for Biomanufacturing Chun You and Y.-H. Percival Zhang Abstract Although cell-free biosystems have been used as a tool for investigating fundamental aspects of biological systems for more than 100 years, they are becoming an emerging biomanufacturing platform in the production of low-value biocommodities (e.g., H2, ethanol, and isobutanol), fine chemicals, and high-value protein and carbohydrate drugs and their precursors. Here we would like to define the cell-free biosystems containing more than three catalytic components in a single reaction vessel, which although different from one-, two-, or three-enzyme biocatalysis can be regarded as a straightforward extension of multienzymatic biocatalysis. In this chapter, we compare the advantages and disadvantages of cell-free biosystems versus living organisms, briefly review the history of cell-free biosystems, highlight a few examples, analyze any remaining obstacles to the scale-up of cell-free biosystems, and suggest potential solutions. Cell-free biosystems could become a disruptive technology to microbial fermentation, especially in the production of high-impact low-value biocommodities mainly due to the very high product yields and potentially low production costs.
Keywords Biocommodity engineering Bioeconomy Biofuels Biomanufacturing Cascade enzyme biocatalyst Cell-free synthetic biology Synthetic pathway biotransformation
Revised book chapter for Advances in Biochemical Engineering/Biotechnology. Volume: Future Trends in Biotechnology. Volume editor: Jian-Jiang Zhong. C. You Y.-H. P. Zhang (&) Biological Systems Engineering Department, Virginia Tech, 304 Seitz Hall, Blacksburg, VA 24061, USA e-mail: [email protected] Y.-H. P. Zhang Institute for Critical Technology and Applied Science (ICTAS), Virginia Tech, Blacksburg, VA 24061, USA Y.-H. P. Zhang Gate Fuels Inc., 2200 Kraft Drive, Suite 1200B, Blacksburg, VA 24060, USA
C. You and Y.-H. Percival Zhang
Contents 1 2 3
Introduction.............................................................................................................................. History of Cell-Free Biosystems............................................................................................. Examples of Cell-Free Biosystems for Biomanufacturing .................................................... 3.1 Cell-Free Biosystems for Biocommodity Engineering ................................................. 3.2 Cell-Free Biosystems for High-Value Product Synthesis ............................................. 3.3 Cell-Free Protein Synthesis ............................................................................................ 4 Challenges................................................................................................................................ 4.1 Low-Cost Enzyme Production and Purification ............................................................ 4.2 Prolonging Enzyme Lifetime .......................................................................................
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