Division-of-labor Among Multicellular System And Applications

The rapid development of synthetic biology from modules,circuits,metabolic pathways to network had greatly expanded the biofunctions in mono-culture.As the breadth and depth of the design continued to increase,more and more multicellular collaboration in microorganism were used in complex biosynthesis processes,with the advantages of reducing burden in mono-culture and flexible dynamic regulation.However,how to rational division-of-labor in multicellular collaboration is still a key problem to be solved.Under the guidance of the "design-build-test-learn" cycle of synthetic biology,based on the principle of "division-of-labor",we constructed a threespecies microbial consortium for power generation and synthesized a 1.14 Mb complex genome based on the exchange of metabolites or the transfer of genetic materials among multicellular collaboration.Synthetic microbial system for bioelectricity can convert chemical energy into electrical energy,which has wide popularization and potential applications.Shewanella oneidensis as a model exoelectrogen has limitations of low energy conversion efficiency and finite carbon source spectrum.To solve these problems,we introduced the high-yielding lactate producing engineered Escherichia coli and the high-riboflavin producing engineered Bacillus subtilis into the system step by step to construct a threespecies microbial consortium for power generation.In this consortium,the three species formed a cross-talk microbial consortium,which performed ‘‘better together’’ on productivity,stability and robustness.As a result,glucose(11 m M,total 0.28 g)was converted to electricity for more than 15 days with high energy conversion efficiency(up to 55.7%),which is the highest level for S.oneidensis.DNA synthesis is the basic and fundamental technology of synthetic biology.The development of synthetic genomics will accelerate the depth expansion of genome design.Traditional methods for assembling synthetic large mammalian genome DNA in one yeast cell was quite difficult due to the sequence was relatively complex and contained a large number of repetitive elements.Here,based on the principle of division-of-labor,the 1.14 Mb genome was split into 4 large DNA fragments: SynA(330 kb)、SynG(268 kb)、SynB(276 kb)and SynC(268 kb),which were assembled in four different yeast cells with different mating types α and a.Based on the transfer of genetic materials by mating and fusion of yeast,four large DNA fragments were assembled in vivo by two rounds DNA splicing and finally assembled into a complete1.14 Mb genome.And further study was the exploration of interactions between the chassis S.cerevisiae and the synthetic heterologous genome through multi-omics analysis.In summary,we firstly constructed a three-species microbial consortium for power generation and synthesized a 1.14 Mb mammalian genome in yeast based on the principle of division-of-labor.Depend on the transfer of genetic material and the exchange of metabolites,we explored the functional allocation strategy among multiple cells.This study provides a new complete division-of-labor strategy for designing more complicated and efficient of multicellular collaboration applied to complex synthetic biological processes.Besides,the allocation strategy could also enhance our understanding of complex microbial consortia in natural environments.