A synthetic biochemistry module for production of bio-based chemicals from glucose

Abstract

Synthetic biochemistry, the cell-free production of biologically based chemicals, is a potentially high-yield, flexible alternative to in vivo metabolic engineering. To limit costs, cell-free systems must be designed to operate continuously with minimal addition of feedstock chemicals. We describe a robust, efficient synthetic glucose breakdown pathway and implement it for the production of bioplastic. The system's performance suggests that synthetic biochemistry has the potential to become a viable industrial alternative.

High-Throughput Screening of Coenzyme Preference Change of Thermophilic 6-Phosphogluconate Dehydrogenase from NADP+ to NAD+

“…The replacement to the other acidic amino acid glutamate at the same position was also found at A30E/R31I/T32D with 3-fold lower K m for NAD + as compared to R31I. Recently, the mutant included replacement to aspartate at same position was reported for 6PGDH from G. stearothermophilus with slightly decreased K m 46. The mutation threonine to isoleucine at position 32 broke the residual hydrogen bonds with 2′-phosphate of NADP + and possibly decreased enzyme binding with NADP + , another 55-fold decrease in catalytic efficiency for NADP + .…”

A cell-free synthetic biochemistry platform for raspberry ketone production

“…Using raspberry ketone as a model pathway, we detail a purified enzyme approach to achieve a high-yield synthesis in a one-pot reaction. Together with other recent synthetic biochemistry studies 11,13 , we feel this approach is potentially expandable to other high-value fine chemicals that require cost efficient cofactor regeneration (i.e. malonyl-CoA, ATP) for in vitro synthesis.…”

“…For fine chemical biomanufacturing, synthetic biology aims to provide green solutions to replacing traditional petroleum or arable farming based production methods 1–6 . The synthetic biochemistry approach, whereby metabolic pathways can be entirely reconstituted within a test-tube with purified enzymes 7–11 or cell-free extracts 12–14 , offers a realistic concept to traditional cell based engineering, with the potential for high performance synthesis of fine chemicals and recombinant proteins to the industrial scale 11,15 .…”

An in vitro synthetic biology platform for the industrial biomanufacturing of myo‐inositol from starch

“…The in vitro (cell-free) synthetic biology platform comprised of numerous (i.e., more than four) purified enzymes in one vessel or called systems biocatalysis has been proposed to become an emerging biomanufacturing platform (Bujara et al, 2011;Fessner, 2015;Fessner and Walter, 1992;Guterl et al, 2012;Hodgman and Jewett, 2012;Hold et al, 2016;Krutsakorn et al, 2013;Opgenorth et al, 2016;Pardee et al, 2016;Rollin et al, 2015;Tessaro et al, 2015;Zhang et al, 2017;Zhu et al, 2014). Beyond whole-cell constraints, such as, cell membrane, bioenergetics, complicated regulation, and so on, this biomanufacturing platform features high product yields (Opgenorth et al, 2016;Rollin et al, 2015;Zhu et al, 2014), fast reaction or biomanufacturing rates Pardee et al, 2016;Zhu and Zhang, 2017;Zhu et al, 2014), easy process control and optimization (Hold et al, 2016;Opgenorth et al, 2016;Zhu and Zhang, 2017), biosystem robustness (Hodgman and Jewett, 2012;Zhu and Zhang, 2017), great biosystem flexibility (Zhu and Zhang, 2017), easy product separation Satoh et al, 2003), implementation of non-natural reactions and pathways You et al, 2013), and so on. However, there are no commercial production examples based on this platform.…”