Culture-free identification of fast-growing cyanobacteria cells by Raman-activated gravity-driven encapsulation and sequencing  

作　　者：Jinyu Cui Rongze Chen Huili Sun Yingyi Xue Zhidian Diao Jingyun Song Xiaohang Wang Jia Zhang Chen Wang Bo Ma Jian Xu Guodong Luan Xuefeng Lu 

机构地区：[1]Key Laboratory of Biofuels,Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Sciences,No.189 Songling Road,Qingdao,266101,China [2]Shandong Energy Institute,Qingdao,266101,China [3]Qingdao New Energy Shandong Laboratory,Qingdao,266101,China [4]College of Life Science,University of Chinese Academy of Sciences,Beijing,China [5]Dalian National Laboratory for Clean Energy,Dalian,116023,China [6]Laboratory for Marine Biology and Biotechnology,Qingdao National Laboratory for Marine Science and Technology,Qingdao,China

出　　处：《Synthetic and Systems Biotechnology》2023年第4期708-715,共8页合成和系统生物技术（英文）

基　　金：supported by the National Key Research and Development Program of China(Grant number 2021YFA0909700);the National Natural Science Foundation of China(Grant numbers 32070084,32270103,32271484,32300058);the Youth Innovation Promotion Association CAS(to Guodong Luan);Postdoctoral Innovation Project of Shandong Province(Grant number SDCX-ZG-202202036);Postdoctoral Science Foundation of China(Grant number 2021M703320);the Shandong Taishan Scholarship(to Xuefeng Lu,and to Guodong Luan).

摘　　要：By directly converting solar energy and carbon dioxide into biobased products,cyanobacteria are promising chassis for photosynthetic biosynthesis.To make cyanobacterial photosynthetic biosynthesis technology economically feasible on industrial scales,exploring and engineering cyanobacterial chassis and cell factories with fast growth rates and carbon fixation activities facing environmental stresses are of great significance.To simplify and accelerate the screening for fast-growing cyanobacteria strains,a method called Individual Cyanobacteria Vitality Tests and Screening(iCyanVS)was established.We show that the ^(13)C incorporation ratio of carotenoids can be used to measure differences in cell growth and carbon fixation rates in individual cyanobacterial cells of distinct genotypes that differ in growth rates in bulk cultivations,thus greatly accelerating the process screening for fastest-growing cells.The feasibility of this approach is further demonstrated by phenotypically and then genotypically identifying individual cyanobacterial cells with higher salt tolerance from an artificial mutant library via Raman-activated gravity-driven encapsulation and sequencing.Therefore,this method should find broad applications in growth rate or carbon intake rate based screening of cyanobacteria and other photosynthetic cell factories.

关 键 词：CYANOBACTERIA Single-cell Raman microspectroscopy Stable isotope probing Carotenoids IDENTIFICATION 

分 类 号：Q813[生物学—生物工程]