Cellulose is the most abundant renewable resource on earth and can be used to produce biofuels and bio-based chemicals. Compared with the traditional microbial fermentation method using cellulose for biological manufacturing, the in vitro multi-enzyme system has strong operability, high product yield, and fast reaction speed, and has been successfully applied to catalyze the complete conversion of cellulose to inositol. However, in the in vitro multi-enzyme pathway using cellulose to generate electricity or hydrogen, due to the high activation energy of the reaction pathway, the low specific enzyme activity of the key enzyme, and poor downstream reaction pulling ability, the initial reaction speed and conversion efficiency of the entire reaction system remained restricted.
Recently, the research team led by You Chun and Zhu Zhiguang, researchers of the Center for In vitro Synthetic Biology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, has improved the overall reaction system by constructing a multi-enzyme complex in a cellulase in vitro multi-enzyme system Power generation efficiency. Based on the pre-designed enzymatic phospholysis pathway that uses an in vitro multi-enzyme system to stoichiometrically convert cellulose biomass, the team combined the rate-limiting enzyme (cellooligosaccharide phosphorylase) and downstream cascade enzymes Glucose phosphate mutase) is constructed as different types of multi-enzyme complexes (Figure a), in which the optimal multi-enzyme complex catalyzes the reaction rate of phospholysis of cellulose to glucose 6-phosphate, which can be increased by a factor of 2.4. The enzyme complex is used in a multi-enzyme system for cellulosic electricity generation. Compared to the free enzyme system, the current density and power density are increased by 3.35 times and 2.14 times, respectively (Figure b, c). This study first reported the use of cellulose to produce bioelectricity, and provided a new idea for improving the initial reaction speed and conversion efficiency of an in vitro multi-enzyme system.
The research was supported by the key deployment projects of the Chinese Academy of Sciences and the National Natural Science Foundation of China, and related research results have been published in Biotechnology for Biofuels. Assistant Researcher Meng Dongdong of Tianjin Institute of Workers and Students is the first author of the paper, and You Chun and Zhu Zhiguang are co-corresponding authors of the paper.
By constructing a multi-enzyme complex, the efficiency of in vitro multi-enzyme system to catalyze the biological production of cellulose is improved. (A) An in vitro multi-enzyme system that catalyzes the production of biofuel from cellulose; (b) The free enzyme system has poor ability to catalyze the production of glucose 6-phosphate from cellulose; (c) The multi-enzyme complex promotes the decomposition of cellulose phosphate to produce glucose 6 -Phosphoric acid.
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