

浏览全部资源
扫码关注微信
1.中粮营养健康研究院有限公司,北京 102209
2.营养健康与食品安全北京市重点实验室,北京 102209
Received:17 March 2023,
Revised:2023-05-08,
Published:31 October 2023
移动端阅览
赵国淼, 杨鑫, 张媛, 王靖, 谭剑, 魏超, 周娜娜, 李凡, 王小艳. 生物设施平台及其工业应用[J]. 合成生物学, 2023, 4(5): 892-903
ZHAO Guomiao, YANG Xin, ZHANG Yuan, WANG Jing, TAN Jian, WEI Chao, ZHOU Nana, LI Fan, WANG Xiaoyan. Biofoundry and its industrial application[J]. Synthetic Biology Journal, 2023, 4(5): 892-903
赵国淼, 杨鑫, 张媛, 王靖, 谭剑, 魏超, 周娜娜, 李凡, 王小艳. 生物设施平台及其工业应用[J]. 合成生物学, 2023, 4(5): 892-903 DOI: 10.12211/2096-8280.2023-024.
ZHAO Guomiao, YANG Xin, ZHANG Yuan, WANG Jing, TAN Jian, WEI Chao, ZHOU Nana, LI Fan, WANG Xiaoyan. Biofoundry and its industrial application[J]. Synthetic Biology Journal, 2023, 4(5): 892-903 DOI: 10.12211/2096-8280.2023-024.
传统菌株改造和筛选实验存在操作烦琐、耗时、易错、难以规模化等问题,生物设施平台将自动化、机器人技术、数据分析与生物研究相结合,通过导轨和机械手臂实现自动化操作,提高了实验操作的稳定性,通过缩小培养体积(微孔板或微液滴),提高了培养和筛选通量,解决了上述问题,大大提高了研发效率。本文简单介绍了自动化设施平台的发展和常见的高通量检测方法,重点介绍了中粮营养健康研究院的自动化设施平台,并结合开展的项目叙述了平台在生物燃料菌株开发、传统酿造菌株筛选、酶的定向进化和筛选等领域的应用,可以预见自动化和高通量化在菌株改造和筛选方向巨大的应用价值。实验室自动化是涉及机械工程、自动化、计算机和生命科学等学科的交叉领域,需要各方面共同努力,才能推动实验室向更高程度的自动化和智能化方向发展。
Traditional methods for strain isolation and improvement can be cumbersome
time-consuming
error-prone
and difficult to scale up
leading to inefficiencies in research and development workflows. With the integration of automation and robotics technology
biofoundry can achieve automated operations through guide rails and robotic arms
leading to improved stability and precision of experimental operations. Additionally
by utilizing smaller cultivation volumes
such as microplates or droplets
the cultivation and screening throughput can be increased
addressing the currently existing issues of traditional methods. This can greatly improve research and development efficiency
allowing for the testing and optimization of large numbers of microbial strains or genetic variants in a high-throughput manner. The biofoundry encompasses interdisciplinary fields such as mechanical engineering
automation
computer science
and life sciences. The collaboration among these fields is crucial for the development and advancement of laboratory automation. By leveraging automation and high-throughput technologies
the field of strain isolation and improvement can benefit from increased efficiency
improved reliability
and scalability. These advancements can accelerate the progress of microbial strain engineering for various applications in biotechnology
medicine
agriculture
and energy production. This paper briefly introduces the composition and classification of the biofoundry
the high-throughput detection method
and focuses on the high-throughput screening platform built by the research team from Nutrition & Health Research Institute
COFCO. In combination with the projects carried out
it introduces the application of the high-throughput screening platform in the fields of biofuel strain development
traditional brewing strain screening
feed substitute antimicrobial screening
directed evolution and screening of enzymes
etc.
2
LEE S Y , KIM H U , CHAE T U , et al . A comprehensive metabolic map for production of bio-based chemicals [J ] . Nature Catalysis , 2019 , 2 ( 1 ): 18 - 33 .
CHEN X L , GAO C , GUO L , et al . DCEO biotechnology: tools to design, construct, evaluate, and optimize the metabolic pathway for biosynthesis of chemicals [J ] . Chemical Reviews , 2018 , 118 ( 1 ): 4 - 72 .
RUGBJERG P , SOMMER M O A . Overcoming genetic heterogeneity in industrial fermentations [J ] . Nature Biotechnology , 2019 , 37 ( 8 ): 869 - 876 .
WEHRS M , TANJORE D , ENG T , et al . Engineering robust production microbes for large-scale cultivation [J ] . Trends in Microbiology , 2019 , 27 ( 6 ): 524 - 537 .
LEE S Y , KIM H U . Systems strategies for developing industrial microbial strains [J ] . Nature Biotechnology , 2015 , 33 ( 10 ): 1061 - 1072 .
ZHANG X , ZHANG X M , XU G Q , et al . Integration of ARTP mutagenesis with biosensor-mediated high-throughput screening to improve L-serine yield in Corynebacterium glutamicum [J ] . Applied Microbiology and Biotechnology , 2018 , 102 ( 14 ): 5939 - 5951 .
GU Y , XU X H , WU Y K , et al . Advances and prospects of Bacillus subtilis cellular factories: from rational design to industrial applications [J ] . Metabolic Engineering , 2018 , 50 : 109 - 121 .
CARBONELL P , JERVIS A J , ROBINSON C J , et al . An automated Design-Build-Test-Learn pipeline for enhanced microbial production of fine chemicals [J ] . Communications Biology , 2018 , 1 : 66 .
ZENG W Z , GUO L K , XU S , et al . High-throughput screening technology in industrial biotechnology [J ] . Trends in Biotechnology , 2020 , 38 ( 8 ): 888 - 906 .
QUAGLIA D , EBERT M C C J C , MUGFORD P F , et al . Enzyme engineering: a synthetic biology approach for more effective library generation and automated high-throughput screening [J ] . PLoS One , 2017 , 12 ( 2 ): e0171741 .
ZHANG Y V , ROCKWOOD A . Impact of automation on mass spectrometry [J ] . Clinica Chimica Acta , 2015 , 450 : 298 - 303 .
LONGWELL C K , LABANIEH L , COCHRAN J R . High-throughput screening technologies for enzyme engineering [J ] . Current Opinion in Biotechnology , 2017 , 48 : 196 - 202 .
RAN C , MISHRA S , TONG S , et al . Engineering biological systems using automated biofoundries [J ] . Metabolic Engineering , 2017 , 42 : 98 - 108 .
ZHANG J Z , CHEN Y C , FU L H , et al . Accelerating strain engineering in biofuel research via build and test automation of synthetic biology [J ] . Current Opinion in Biotechnology , 2021 , 67 : 88 - 98 .
LE FEUVRE R A , SCRUTTON N S . A living foundry for Synthetic Biological Materials: a synthetic biology roadmap to new advanced materials [J ] . Synthetic & Systems Biotechnology , 2018 , 3 ( 2 ): 105 - 112 .
HILLSON N , CADDICK M , CAI Y Z , et al . Building a global alliance of biofoundries [J ] . Nature Communications , 2019 , 10 : 2040 .
ENGHIAD B , XUE P , SINGH N , et al . PlasmidMaker is a versatile, automated, and high throughput end-to-end platform for plasmid construction [J ] . Nature Communications , 2022 , 13 : 2697 .
ZHAO H M . Illinois biological foundry for advanced biomanufacturing (iBioFAB) [C/OL ] . Synthetic Biology: Engineering , Evolution , and Design Conference 2015 , SEED 2015, 2015 , 2: 784 - 785 [ 2023-03-01 ] . https://experts.illinois.edu/en/publications/illinois-biological-foundry-for-advanced-biomanufacturing-ibiofab https://experts.illinois.edu/en/publications/illinois-biological-foundry-for-advanced-biomanufacturing-ibiofab .
HAMEDIRAD M , CHAO R , WEISBERG S , et al . Towards a fully automated algorithm driven platform for biosystems design [J ] . Nature Communications , 2019 , 10 : 5150 .
XU K , QIN L , BAI W X , et al . Multilevel defense system (MDS) relieves multiple stresses for economically boosting ethanol production of industrial Saccharomyces cerevisiae [J ] . ACS Energy Letters , 2020 , 5 ( 2 ): 572 - 582 .
LIU W S , JIANG R R . Combinatorial and high-throughput screening approaches for strain engineering [J ] . Applied Microbiology and Biotechnology , 2015 , 99 ( 5 ): 2093 - 2104 .
ALPER H , MIYAOKU K , STEPHANOPOULOS G . Construction of lycopene-overproducing E. coli strains by combining systematic and combinatorial gene knockout targets [J ] . Nature Biotechnology , 2005 , 23 ( 5 ): 612 - 616 .
ÖZAYDIN B , BURD H , LEE T S , et al . Carotenoid-based phenotypic screen of the yeast deletion collection reveals new genes with roles in isoprenoid production [J ] . Metabolic Engineering , 2013 , 15 : 174 - 183 .
ZELCBUCH L , ANTONOVSKY N , BAR-EVEN A , et al . Spanning high-dimensional expression space using ribosome-binding site combinatorics [J ] . Nucleic Acids Research , 2013 , 41 ( 9 ): e98 .
LEE J H , LEE S H , YIM S S , et al . Quantified high-throughput screening of Escherichia coli producing poly(3-hydroxybutyrate) based on FACS [J ] . Applied Biochemistry and Biotechnology , 2013 , 170 ( 7 ): 1767 - 1779 .
TYO K E J , JIN Y S , ESPINOZA F A , et al . Identification of gene disruptions for increased poly-3-hydroxybutyrate accumulation in Synechocystis PCC 6803 [J ] . Biotechnology Progress , 2009 , 25 ( 5 ): 1236 - 1243 .
KLEIN-MARCUSCHAMER D , SANTOS C N S , YU H M , et al . Mutagenesis of the bacterial RNA polymerase alpha subunit for improvement of complex phenotypes [J ] . Applied and Environmental Microbiology , 2009 , 75 ( 9 ): 2705 - 2711 .
ALPER H , MOXLEY J , NEVOIGT E , et al . Engineering yeast transcription machinery for improved ethanol tolerance and production [J ] . Science , 2006 , 314 ( 5805 ): 1565 - 1568 .
BASAK S , GENG H F , JIANG R R . Rewiring global regulator cAMP receptor protein (CRP) to improve E. coli tolerance towards low pH [J ] . Journal of Biotechnology , 2014 , 173 : 68 - 75 .
LIU H M , YAN M , LAI C G , et al . gTME for improved xylose fermentation of Saccharomyces cerevisiae [J ] . Applied Biochemistry and Biotechnology , 2010 , 160 ( 2 ): 574 - 582 .
CHONG H Q , HUANG L , YEOW J , et al . Improving ethanol tolerance of Escherichia coli by rewiring its global regulator cAMP receptor protein (CRP) [J ] . PLoS One , 2013 , 8 ( 2 ): e57628 .
HENNING H , LEGGEWIE C , POHL M , et al . Identification of novel benzoylformate decarboxylases by growth selection [J ] . Applied and Environmental Microbiology , 2006 , 72 ( 12 ): 7510 - 7517 .
PFLEGER B F , PITERA D J , SMOLKE C D , et al . Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes [J ] . Nature Biotechnology , 2006 , 24 ( 8 ): 1027 - 1032 .
BOERSMA Y L , DRÖGE M J , VAN DER SLOOT A M , et al . A novel genetic selection system for improved enantioselectivity of Bacillus subtilis lipase A [J ] . ChemBioChem , 2008 , 9 ( 7 ): 1110 - 1115 .
BOLES E , OREB M . A growth-based screening system for hexose transporters in yeast [M/OL ] // Methods in Molecular Biology . New York, NY: Springer New York, 2018 , 1713 : 123 - 135 [2023-03-01] . https://link.springer.com/protocol/10.1007/978-1-4939-7507-5_10 https://link.springer.com/protocol/10.1007/978-1-4939-7507-5_10 .
DIETRICH J A , MCKEE A E , KEASLING J D . High-throughput metabolic engineering: advances in small-molecule screening and selection [J ] . Annual Review of Biochemistry , 2010 , 79 : 563 - 590 .
LATCHMAN D S . Transcription factors: an overview [J ] . The International Journal of Biochemistry & Cell Biology , 1997 , 29 ( 12 ): 1305 - 1312 .
BINDER S , SCHENDZIELORZ G , STÄBLER N , et al . A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level [J ] . Genome Biology , 2012 , 13 ( 5 ): R40 .
MAHR R , GÄTGENS C , GÄTGENS J , et al . Biosensor-driven adaptive laboratory evolution of l-valine production in Corynebacterium glutamicum [J ] . Metabolic Engineering , 2015 , 32 : 184 - 194 .
BASTET L , TURCOTTE P , WADE J T , et al . Maestro of regulation: riboswitches orchestrate gene expression at the levels of translation, transcription and mRNA decay [J ] . RNA Biology , 2018 : 15 ( 6 ): 679 - 682 .
ECKDAHL T T , CAMPBELL A M , HEYER L J , et al . Programmed evolution for optimization of orthogonal metabolic output in bacteria [J ] . PLoS One , 2015 , 10 ( 2 ): e0118322 .
DIXON N , DUNCAN J N , GEERLINGS T , et al . Reengineering orthogonally selective riboswitches [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2010 , 107 ( 7 ): 2830 - 2835 .
WANG J L , WEI J H , SU S H , et al . Novel fluorescence resonance energy transfer optical sensors for vitamin B 12 detection using thermally reduced carbon dots [J ] . New Journal of Chemistry , 2015 , 39 ( 1 ): 501 - 507 .
NGUYEN T T T , HUY B T , TAWFIK S M , et al . Highly selective and sensitive optosensing of glutathione based on fluorescence resonance energy transfer of upconversion nanoparticles coated with a Rhodamine B derivative [J ] . Arabian Journal of Chemistry , 2020 , 13 ( 1 ): 2671 - 2679 .
DING Y D , LI J , ENTERINA J R , et al . Ratiometric biosensors based on dimerization-dependent fluorescent protein exchange [J ] . Nature Methods , 2015 , 12 ( 3 ): 195 - 198 .
FU X Z , ZHANG Y Y , XU Q , et al . Recent advances on sorting methods of high-throughput droplet-based microfluidics in enzyme directed evolution [J ] . Frontiers in Chemistry , 2021 , 9 : 666867 .
ZHANG Z D , GUO Q , WANG Y T , et al . High-throughput screening of microbial strains in large-scale microfluidic droplets [J ] . Frontiers in Bioengineering and Biotechnology , 2023 , 11 : 1105277 .
UTHARALA R , GRAB A , VAFAIZADEH V , et al . A microfluidic Braille valve platform for on-demand production, combinatorial screening and sorting of chemically distinct droplets [J ] . Nature Protocols , 2022 , 17 ( 12 ): 2920 - 2965 .
KÖRFER G , PITZLER C , VOJCIC L , et al . In vitro flow cytometry-based screening platform for cellulase engineering [J ] . Scientific Reports , 2016 , 6 : 26128 .
WANG Y T , ZHANG X X , SHANG L R , et al . Thriving microfluidic technology [J ] . Science Bulletin , 2021 , 66 ( 1 ): 9 - 12 .
涂然 , 李世新 , 李昊霓 , 等 . 液滴微流控技术在微生物工程菌株选育中的应用进展 [J ] . 合成生物学 , 2023 ( 1 ): 165 - 184 .
TU R , LI S X , LI H N , et al . Advances and applications of droplet-based microfluidics in evolution and screening of engineered microbial strains [J ] . Synthetic Biology Journal , 2023 ( 1 ): 165 - 184 .
LEAMON J H , LINK D R , EGHOLM M , et al . Overview: methods and applications for droplet compartmentalization of biology [J ] . Nature Methods , 2006 , 3 ( 7 ): 541 - 543 .
GIELEN F , HOURS R , EMOND S , et al . Ultrahigh-throughput-directed enzyme evolution by absorbance-activated droplet sorting (AADS) [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2016 , 113 ( 47 ): E7383 - E7389 .
BARET J C , MILLER O J , TALY V , et al . Fluorescence-activated droplet sorting (FADS): efficient microfluidic cell sorting based on enzymatic activity [J ] . Lab on a Chip , 2009 , 9 ( 13 ): 1850 - 1858 .
GOTO H , KANAI Y , YOTSUI A , et al . Microfluidic screening system based on boron-doped diamond electrodes and dielectrophoretic sorting for directed evolution of NAD(P)-dependent oxidoreductases [J ] . Lab on a Chip , 2020 , 20 ( 4 ): 852 - 861 .
WANG X X , REN L H , SU Y T , et al . Raman-activated droplet sorting (RADS) for label-free high-throughput screening of microalgal single-cells [J ] . Analytical Chemistry , 2017 , 89 ( 22 ): 12569 - 12577 .
SESEN M , WHYTE G . Image-based single cell sorting automation in droplet microfluidics [J ] . Scientific Reports , 2020 , 10 : 8736 .
HOLLAND-MORITZ D A , WISMER M K , MANN B F , et al . Mass activated droplet sorting (MADS) enables high-throughput screening of enzymatic reactions at nanoliter scale [J ] . Angewandte Chemie International Edition , 2020 , 59 ( 11 ): 4470 - 4477 .
王小艳 , 秦磊 , 刘辉 , 等 . 淀粉质燃料乙醇发酵胁迫及菌株耐受性改造 [J ] . 精细化工 , 2019 , 36 ( 4 ): 568 - 574 .
WANG X Y , QIN L , LIU H , et al . Research progress of starchy fuel ethanol fermentation and the tolerance of Saccharomyces cerevisiae [J ] . Fine Chemicals , 2019 , 36 ( 4 ): 568 - 574 .
PHAN A P H , NGO T T , LENHOFF H M . Spectrophotometric assay for lysine decarylase [J ] . Analytical Biochemistry , 1982 , 120 ( 1 ): 193 - 197 .
夏冰 , 丁子元 , 郑晓卫 , 等 . 植物乳杆菌和菌剂及其在生物胺降解、黄酒生产中的应用 : CN111254101B [P ] . 2020-07-28 .
XIA B , DING Z Y , ZHENG X W , et al . Lactobacillus plantarum, microbial inoculum and application of Lactobacillus plantarum and microbial inoculum in biogenic amine degradation and yellow rice wine production : CN111254101B [P ] . 2020-07-28 .
王德昌 , 明明 , 周维广 . 分光光度法测定高级醇 [J ] . 啤酒科技 , 2005 ( 3 ): 37 - 38 .
WANG D C , MING M , ZHOU W G . Spectrophotometric determination of higher alcohols [J ] . Beer Science and Technology , 2005 ( 3 ): 37 - 38 .
杨鑫 , 孙浩轩 , 何伟 , 等 . 一步法制备四甲基吡嗪用反应装置 : CN218077901U [P ] . 2022-12-20 .
YANG X , SUN H X , HE W , et al . Reaction device for preparing tetramethylpyrazine by one-step method : CN218077901U [P ] . 2022-12-20 .
丁子元 , 杨鑫 , 靳喜庆 , 等 . 枯草芽孢杆菌、菌剂、应用及制备四甲基吡嗪的方法 : CN115386525B [P ] . 2023-01-31 .
DING Z Y , YANG X , JIN X Q , et al . Bacillus subtilis , fungicide , application and method for preparing tetramethylpyrazine: CN115386525B [P ] . 2023-01-31 .
KONG C L , LI A H , SU J , et al . Flavor modification of dry red wine from Chinese spine grape by mixed fermentation with Pichia fermentans and S. cerevisiae [J ] . LWT , 2019 , 109 : 83 - 92 .
刘沛通 , 丁子元 , 于庆泉 , 等 . 优良本土酿酒酵母的酿造特性研究 [J ] . 中国食品学报 , 2023 , 23 ( 1 ), 204 - 215 .
LIU P T , DING Z Y , YU Q Q , et al . Studies on oenological characteristics of high-quality Chinese indigenous Saccharomyces cerevisiae strains [J ] . Journal of Chinese Institute of Food Science and Technology , 2023 , 23 ( 1 ), 204 - 215 .
刘沛通 , 丁子元 , 郑晓卫 , 等 . 酿酒酵母和菌剂以及它们在制备发酵产品特别是怀涿盆地葡萄酒酿造中的应用 : CN111961603B [P ] . 2021-01-01 .
LIU P T , DING Z Y , ZHENG X W , et al . Saccharomyces cerevisiae and microbial agent as well as application thereof to preparation of fermented product and particularly brewing of wine in Huazhuo Basin : CN111961603B [P ] . 2021-01-01 .
郑晓卫 , 刘沛通 , 李泽福 , 等 . 抗逆性优良的空间育种酿酒酵母及其应用 : CN115651852B [P ] . 2023-04-11 .
ZHENG X W , LIU P T , LI Z F , et al . Spatial breeding Saccharomyces cerevisiae with excellent stress resistance and application thereof : CN115651852B [P ] . 2023-04-11 .
王千 , 白杰 , 江会锋 . 合成生物学酶改造设计技术的研究进展 [J ] . 生命科学 , 2021 , 33 ( 12 ): 1493 - 1501 .
WANG Q , BAI J , JIANG H F . Research progress on technologies of enzyme engineering and design in synthetic biology [J ] . Chinese Bulletin of Life Sciences , 2021 , 33 ( 12 ): 1493 - 1501 .
赵聪敏 . 甜菊糖苷单体分离、甜味特性及应用研究 [D ] . 邯郸 : 河北工程大学 , 2021 .
ZHAO C M . Isolation of stevia glycoside, sweetness characteristics and application [D ] . Handan : Hebei University of Engineering , 2021 .
祁飞 , 刘瑞敏 , 张真真 . 一种通过易错PCR技术及高通量筛选提高葡萄糖基转移酶EUGT 11 酶活方法 : CN113584016A [P ] . 2021-11-02 .
QI F , LIU R M , ZHANG Z Z . Method for improving enzyme activity of glucosyltransferase EUGT 11 through error-prone PCR technology and high-throughput screening : CN113584016A [P ] . 2021-11-02 .
谭剑 , 佟毅 , 赵国淼 , 等 . 玉米赤霉烯酮浓度及其降解酶酶活力的测定方法以及玉米赤霉烯酮降解菌的筛选方法 : CN112577930A [P ] . 2021-03-30 .
TAN J , TONG Y , ZHAO G M , et al . Zearalenone concentration and zearalenone degrading enzyme activity determination method and zearalenone degrading bacterium screening method : CN112577930A [P ] . 2021-03-30 .
赵国淼 , 佟毅 , 谭剑 , 等 . 测定呕吐毒素浓度的方法及其应用 : CN114813895A [P ] . 2022-07-29 .
ZHAO G M , TONG Y , TAN J , et al . Method for determining vomitoxin concentration and application thereof : CN114813895A [P ] . 2022-07-29 .
0
Views
1
下载量
0
CSCD
Publicity Resources
Related Articles
Related Author
Related Institution
京公网安备11010802024621