近年来，微生物发酵生产技术迭代升级增速成为生物制造产业的重要支撑，随着合成生物学技术快速发展，工业菌株选育能力显著提升。然而，微生物菌种选育的限速环节之一是表型测试，其主要原因在于该环节以摇瓶、孔板、搅拌釜等微型反应器为主，存在溶氧控制困难、搅拌振荡产生气泡多、育种过程设备集成难度大等问题，制约高通量自动化微生物育种技术创新发展。因此，研制新型微型高通量微生物表型筛选与测试反应器系统及其自动化装备是支撑生物产业发展的关键。本文提出并研制了一套新型微型管式微生物育种反应器，结合应用需求对其特性、关键技术及装备集成与应用进行了系统研究，为工业微生物选育和微生物学研究提供微型化、自动化、高通量的研究平台。首先，利用透气性优越的特氟龙材质微管路构建了一套新型微型管式反应器，其在剪切力小于0.2498 Pa的条件下KLa高达600 /h，并为该反应器研制了宽范围（0 - 90%）氧分压的氧传递调控技术和大量程（0 - 20）OD600在线检测技术。在此基础上搭建了具有培养、在线检测功能的微生物培养装置原型机，实现了对6种典型模式微生物的培养，其生长速率和最高OD600值均优于摇瓶培养结果。且通过提高氧分压增大氧传递速率，将枯草芽孢杆菌的最高OD600值从4.68提高至10.34。然后，针对传统微生物适应性进化育种存在通量低，自动化、微型化装备匮乏等问题，研制了基于微型管式反应器的高通量微生物适应性进化装备，并在酿酒酵母耐乙醇适应性进化中成功验证了装备性能，连续培养时间长达240 h，累计传代15次，并将乙醇浓度从0逐渐提高至8%。然后将该装备应用于植物乳杆菌耐高氧适应性进化实验，进化后的耐氧菌株在空气环境下OD600值提升了215.15%。最后，针对现有微生物单克隆挑选方法存在效率低、装备配套多、占地空间大、搭建运行成本高等问题，结合微型管式生物反应器与液滴微流控技术特点，研制了高通量微液滴微生物单克隆挑选装备，并在大肠杆菌的单克隆挑选实验中成功验证了装备性能，所获得菌株的单克隆率为95.65%，和传统固体平板单克隆挑选方法结果一致。然后将其应用于肠道微生物培养组学研究，相比于传统平板分离培养方法，肠道菌群中可培养微生物的数量增加94%，在科水平的种类增加60%。

In recent years, the iterative upgrading of microbial fermentation production technology has become an important support for the development of the biomanufacturing industry, with the rapid development of synthetic biology technology, the ability to breed industrial strains has been significantly improved. However, phenotype testing is the one of the limiting factors in microbial strain breeding. This is mainly due to the fact that this link is mainly based on microreactors such as shake flasks, microplates, and stirred tanks, which are difficult in dissolved oxygen control, produce many bubbles resulting from stirring and oscillation, and have high difficulty in equipment integration. This restricts the innovative development of high-throughput automated microbial breeding technology. Therefore, the development of a novel miniature high-throughput microbial phenotype screening and testing reactor system along with its automated equipment is the key to supporting the development of the biological industry. Here, we propose and develop a set of novel microtubular microbial breeding reactors, and systematically research their characteristics, key technologies and equipment integration, and applications according to the requirements, providing miniaturized, automated, and high-throughput research platforms for industrial microbial breeding and microbiology research.Firstly, a novel type of miniature tubular reactor was constructed based on the Teflon microtube with superior permeability, whose KLa was as high as 600/h under the condition of shear force less than 0.2498 Pa. An oxygen transfer control technology with a wide-range (0-90%) of oxygen partial pressure and a large range (0-20) of OD600 online detection technology were also developed for this reactor. On this basis, a prototype device for microbial culture and on-line detection was constructed, and the culture of 6 typical model microorganisms was realized. It was found that their growth rates and the maximum OD600 values in this device were better than the results of the shake flask culture. At the same time, the OD600 value of Bacillus subtilis was increased from 4.68 to 10.34 in the device by increasing the partial pressure of oxygen to increase the oxygen transfer rate.Next, to address the problems of low throughput and lack of automated and miniaturized equipment in conventional microbial adaptive evolution breeding, a high-throughput microbial adaptive evolution equipment based on microtubular reactor was developed, and the performance of the equipment was successfully verified with the ethanol-tolerant adaptive evolution of Saccharomyces cerevisiae, which was continuously cultured for up to 240 h with 15 times of subculture and gradually increased concentration of ethanol from 0 to 8%. Then the equipment was applied to the oxygen-tolerance adaptive evolution experiment of Lactobacillus plantarum, and the maximum OD600 value of the evolved strain was increased by 215.15% under air environment.Finally, for the existing microbial monoclonal selection methods, there are problems such as low efficiency, large space occupation of equipment, and high cost of construction and operation. To address these problems, a high-throughput microdroplet microbial monoclonal selection equipment was developed in combination with the characteristics of microtubular bioreactor and droplet microfluidic technology. We successfully verified the performance of the equipment in the monoclonal selection experiments of Escherichia coli, and the monoclonal rate of the obtained strains was 95.65%, consistent with the results of the traditional solid plate monoclonal selection method. Then the equipment was applied to the study of gut microbial culturomics. Compared with the solid plate isolation culture method, the number of culturable microorganisms in the gut microbial community increased by 94% and the species at the family level increased by 60%.