In vitro BioTransformation(ivBT):a new frontier of industrial biomanufacturing
Huge challenges,such as food security,energy security,climate change,dual-carbon target,and so on,motivate human society to seek disruptive and innovative solutions.In vitro biotransformation(ivBT),bridging the gap between whole-cell-based fermentation and enzyme-based biocatalysis,is an emerging biomanufacturing platform designed for the production of biocommodities(e.g.,synthetic starch,healthy sweeteners,organic acids,etc.)and bioenergy.In ivBT,in vitro synthetic enzymatic biosystem(ivSEB)is its high-efficiency biocatalyst.Based on the Chinese philosophy that"Tao is simple",ivSEB is the in vitro reconstruction of artificial(non-natural)enzymatic pathways with a number of natural enzymes,artificial enzymes,and/or(biomimetic or natural)coenzymes,and/or artificial membrane,without living cell's constraints,such as cell duplication,bioenergetics,basic metabolisms,regulation,and so on.ivBT enables it to surpass the limitations of whole-cell fermentation and has multiple advantages,such as theoretical product yield,at least 10-time volumetric productivity,tolerance to toxic substrate/product,and so on.This review defines the concept of ivBT,presents its design principles,distinguishes it from other seemingly-like concepts,such as cell-free protein synthesis and cascade enzyme biocatalysis,introduces several representative examples,and discusses its challenges and opportunities.The development of ivBT is based on the linear strategy of"Design-Build-GoNG-Optimization",leading to super-biomanufacturing machines that can meet national needs,such as food security and new energy system.To address food security,we propose two out-of-the-box solutions:(1)in vitro biotransformation of cellulose to starch,possibly increasing the starch supply by a factor of 10;(2)artificial starch synthesis from CO2 by combining ivBT and chemical catalysis.Furthermore,the revolutionary production of starch could open a door to the starch-based carbohydrate economy,wherein starch is a high-density hydrogen carrier,more than 2.5 times that of compressed hydrogen,and an ultra-high electricity storage compound,more than 10 times of lithium-ion battery.In a word,ivBT featuring ultra-high energy efficiency and potentially-low-cost production could become a third industrial biomanufacturing platform and help solve huge challenges.
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