嗜盐菌合成生物学为基础的新型生物制造:生物制造2.0  

New biomanufacturing based on synthetic biology of halophilic bacteria:Biomanufacture 2.0

作　　者：李韶威 何宏韬 吴琼[1,2,3] 陈国强[1,2,3,4] Shaowei Li;Hongtao He;Qiong Wu;Guo-Qiang Chen(School of Life Sciences,Tsinghua University,Beijing 100084,China;Center for Synthetic and Systems Biology,Tsinghua University,Beijing 100084,China;Tsinghua University-Peking University Joint Center for Life Sciences,Tsinghua University,Beijing 100084,China;MOE Key Lab of Industrial Biocatalysis,Department of Chemical Engineering,Tsinghua University,Beijing 100084,China)

机构地区：[1]清华大学生命科学学院,北京100084 [2]清华大学合成与系统生物学中心,北京100084 [3]清华大学-北京大学生命科学联合中心,北京100084 [4]清华大学化工系教育部工业生物催化重点实验室,北京100084

出　　处：《科学通报》2024年第30期4447-4460,共14页Chinese Science Bulletin

基　　金：国家绿色生物制造重大专项(2021YFC2101700);国家合成生物学重大专项(2018YFA0900200)资助。

摘　　要：生物制造是利用生物体机能进行物质加工与合成的绿色生产方式.目前,传统的生物制造以模式微生物为主,具有发酵技术成熟,分子改造技术成熟,但易染菌、能耗高、产率低、过程控制复杂等特点,其生产化工产品的竞争力不如化学工业.为突破传统生物制造的瓶颈,嗜盐菌等极端微生物近10年快速发展成为底盘细胞,成为开放无灭菌以及可以连续发酵的“下一代工业生物技术”(next generation industrial biotechnology,NGIB).嗜盐微生物是在高渗透压环境中具备正常生长能力的极端微生物,近年来,嗜盐菌的分子生物学工具种类不断增加,使得其改造和工程化发展迅速,已经完成了多种聚羟基脂肪酸酯(polyhydroxyalkanoates,PHA)和小分子化合物的生物合成,包括形态学工程改造带来的下游产物回收的便利.基于嗜盐菌底盘细胞的“下一代工业生物技术”具有基因可编辑性、节能节水、无需灭菌、可连续发酵、设备和操作简单以及一个底盘,多种产物等优点,合成生物学武装的嗜盐菌打开了新型生物制造的一扇新窗口,必将引起生物制造的一场新的变革,我们将其命名为“生物制造2.0”.Bioproduction is a green process utilizing living organisms or enzymes to process and synthesize substances and is deemed to have the potential to lead“Low Carbon and Green Manufacturing”.It has attracted increasing attention due to its sustainability.However,traditional bioproduction is mostly based on model microorganisms like E.coli,Bacillus spp.,Saccharomyces cerevisiae,et al.grown under sterile conditions in batch or fed-batch processes,resulting in process complexity,heavy energy consumption for sterilization and higher costs.To overcome the disadvantages,extremophiles such as halophilic bacteria have been exploited as the chassis of the“Next Generation Industrial Biotechnology(NGIB)”allowing open unsterile bioprocessing in seawater.With extremophiles as chassis,bioproduction based on NGIB can avoid sterilization so that it can decrease the cost of water and energy.It shows advantages compared to traditional bioproduction in terms of process complexity and product costs.With the rapid development of molecular tools for halophiles in the last 20 years in this lab,Halomonas spp.can be engineered to produce various polyhydroxyalkanoates(PHA)and many small molecular chemicals as well as proteins together with morphology engineering for convenient downstream processing.For example,gene editing tools like CRISPR/Cas9 and CRISPRi make the construction of exogenous synthetic pathways possible.Additionally,constitutive and inducible promotors make the gene expression tuning reliable.With these tools,the biosynthesis pathway of various PHA monomers and copolymers has been constructed in Halomonas bluephagenesis.The expression tuning of cell size-related genes mreB or ftsZ gene can enlarge bacterial sizes so that they produce more intracellular products during incubations.The morphology engineering of halophiles makes the separation of products easier.Usually,more than half of the cost comes from the separation of products.Longer bacteria are easily settled so centrifugation processes are less demanding.Both o

关 键 词：嗜盐菌 下一代工业生物技术 合成生物学 聚羟基脂肪酸酯 聚羟基丁酸酯 生物制造2.0 

分 类 号：Q81[生物学—生物工程]