Cell-free synthetic enzymatic biosystem is emerging to expand the traditional biotechnological mode by utilizing a number of purified/partially purified enzymes and coenzymes in a single reaction vessel for the produc...Cell-free synthetic enzymatic biosystem is emerging to expand the traditional biotechnological mode by utilizing a number of purified/partially purified enzymes and coenzymes in a single reaction vessel for the production of desired products from low-cost substrates.Here,a cell-free synthetic biosystem containing minimized number of reactions was designed for the conversion of D-glucose to L-lactate via pyruvate.This NADH-balanced biosystem was comprised of only 5 thermophilic enzymes without ATP supplementation.After optimization of enzyme loading amounts,buffer concentration and cofactor concentration,D-glucose was converted to L-lactate with a product yield of∼90%.Our study has provided an emerging platform with potentials in producing pyruvatederived chemicals,and may promote the development of cell-free synthetic enzymatic biosystems for biomanufacturing.展开更多
Although most in vitro(cell-free)synthetic biology projects are usually used for the purposes of fundamental research or the formation of high-value products,in vitro synthetic biology platform,which can implement com...Although most in vitro(cell-free)synthetic biology projects are usually used for the purposes of fundamental research or the formation of high-value products,in vitro synthetic biology platform,which can implement complicated biochemical reactions by the in vitro assembly of numerous enzymes and coenzymes,has been proposed for low-cost biomanufacturing of bioenergy,food,biochemicals,and nutraceuticals.In addition to the most important advantage-high product yield,in vitro synthetic biology platform features several other biomanufacturing advantages,such as fast reaction rate,easy product separation,open process control,broad reaction condition,tolerance to toxic substrates or products,and so on.In this article,we present the basic bottom-up design principles of in vitro synthetic pathway from basic building blocks-BioBricks(thermoenzymes and/or immobilized enzymes)to building modules(e.g.,enzyme complexes or multiple enzymes as a module)with specific functions.With development in thermostable building blocks-BioBricks and modules,the in vitro synthetic biology platform would open a new biomanufacturing age for the cost-competitive production of biocommodities.展开更多
基金the Key Research Program of the Chinese Academy of Sciences(Grant No.ZDRW-ZS-2016-3)National Natural Science Foundation of China(Grant No.31600636).
文摘Cell-free synthetic enzymatic biosystem is emerging to expand the traditional biotechnological mode by utilizing a number of purified/partially purified enzymes and coenzymes in a single reaction vessel for the production of desired products from low-cost substrates.Here,a cell-free synthetic biosystem containing minimized number of reactions was designed for the conversion of D-glucose to L-lactate via pyruvate.This NADH-balanced biosystem was comprised of only 5 thermophilic enzymes without ATP supplementation.After optimization of enzyme loading amounts,buffer concentration and cofactor concentration,D-glucose was converted to L-lactate with a product yield of∼90%.Our study has provided an emerging platform with potentials in producing pyruvatederived chemicals,and may promote the development of cell-free synthetic enzymatic biosystems for biomanufacturing.
基金the National Natural Science Foundation of China(Grant No.31700033)the Key Research Program of the Chinese Academy of Sciences(Grant No.ZDRW-ZS-2016-3).
文摘Although most in vitro(cell-free)synthetic biology projects are usually used for the purposes of fundamental research or the formation of high-value products,in vitro synthetic biology platform,which can implement complicated biochemical reactions by the in vitro assembly of numerous enzymes and coenzymes,has been proposed for low-cost biomanufacturing of bioenergy,food,biochemicals,and nutraceuticals.In addition to the most important advantage-high product yield,in vitro synthetic biology platform features several other biomanufacturing advantages,such as fast reaction rate,easy product separation,open process control,broad reaction condition,tolerance to toxic substrates or products,and so on.In this article,we present the basic bottom-up design principles of in vitro synthetic pathway from basic building blocks-BioBricks(thermoenzymes and/or immobilized enzymes)to building modules(e.g.,enzyme complexes or multiple enzymes as a module)with specific functions.With development in thermostable building blocks-BioBricks and modules,the in vitro synthetic biology platform would open a new biomanufacturing age for the cost-competitive production of biocommodities.
基金supported by the National Key Research and Development Program of China(2022YFA0912300)the Agricultural Science and Technology Innovation Program(ASTIP)Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project,China(TSBICIP-CXRC-067)。
