1. Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Shanghai 200025, China 2. University of the Chinese Academy of Sciences, Beijing 100049, China 3. CAS Key Laboratory of Microbial Physiological and Metebolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China 4. Jiangsu National Synergetic Innovation Center for Advanced Materials, Nanjing Tech University, Nanjing 211816, China
Overproduction of small-molecule chemicals using engineered microbial cells has greatly reduced the production cost and promoted environmental protection. Notably, the rapid and sensitive evaluation of the in vivo concentrations of the desired products greatly facilitates the optimization process of cell factories. For this purpose, many genetic components have been adapted into in vivo biosensors of small molecules, which couple the intracellular concentrations of small molecules to easily detectable readouts such as fluorescence, absorbance, and cell growth. Such biosensors allow a high-throughput screening of the small-molecule products, and can be roughly classified as protein-based and RNA-based biosensors. This review summarizes the recent developments in the design and applications of biosensors for small-molecule products.
Schallmey M, Frunzke J, Eggeling L, Marienhagen J. Looking for the pick of the bunch: High-throughput screening of producing microorganisms with biosensors. Current Opinion in Biotechnology, 2014, 26: 148–154
https://doi.org/10.1016/j.copbio.2014.01.005
2
Ro D K, Paradise E M, Ouellet M, Fisher K J, Newman K L, Ndungu J M, Ho K A, Eachus R A, Ham T S, Kirby J, . Production of the antimalarial drug precursor artemisinic acid in engineered yeast. Nature, 2006, 440(7086): 940–943
https://doi.org/10.1038/nature04640
3
Martin V J J, Pitera D J, Withers S T, Newman J D, Keasling J D. Engineering a mevalonate pathway in Escherichia coli for production of terpenoids. Nature Biotechnology, 2003, 21(7): 796–802
https://doi.org/10.1038/nbt833
Dellomonaco C, Clomburg J M, Miller E N, Gonzalez R. Engineered reversal of the beta-oxidation cycle for the synthesis of fuels and chemicals. Nature, 2011, 476(7360): 355–359
https://doi.org/10.1038/nature10333
6
Enquist-Newman M, Faust A M E, Bravo D D, Santos C N S, Raisner R M, Hanel A, Sarvabhowman P, Le C, Regitsky D D, Cooper S R, . Efficient ethanol production from brown macroalgae sugars by a synthetic yeast platform. Nature, 2013, 505(7482): 239–243
https://doi.org/10.1038/nature12771
7
Becker J, Zelder O, Hafner S, Schroder H, Wittmann C. From zero to hero-design-based systems metabolic engineering of Corynebacterium glutamicum for L-lysine production. Metabolic Engineering, 2011, 13(2): 159–168
https://doi.org/10.1016/j.ymben.2011.01.003
8
Lee K H, Park J H, Kim T Y, Kim H U, Lee S Y. Systems metabolic engineering of Escherichia coli for L-threonine production. Molecular Systems Biology, 2007, 3(1): 149
9
Kind S, Neubauer S, Becker J, Yamamoto M, Volkert M, von Abendroth G, Zelder O, Wittmann C. From zero to hero—production of bio-based nylon from renewable resources using engineered Corynebacterium glutamicum. Metabolic Engineering, 2014, 25: 113–123
https://doi.org/10.1016/j.ymben.2014.05.007
10
Zhang Y X, Perry K, Vinci V A, Powell K, Stemmer W P C, del Cardayre S B. Genome shuffling leads to rapid phenotypic improvement in bacteria. Nature, 2002, 415(6872): 644–646
https://doi.org/10.1038/415644a
11
Wang H H, Isaacs F J, Carr P A, Sun Z Z, Xu G, Forest C R, Church G M. Programming cells by multiplex genome engineering and accelerated evolution. Nature, 2009, 460(7257): 894–898
https://doi.org/10.1038/nature08187
12
Cobb R E, Chao R, Zhao H M. Directed evolution: Past, present, and future. AIChE Journal. American Institute of Chemical Engineers, 2013, 59(5): 1432–1440
https://doi.org/10.1002/aic.13995
13
Alper H, Miyaoku K, Stephanopoulos G. Construction of lycopene-overproducing E. coli strains by combining systematic and combinatorial gene knockout targets. Nature Biotechnology, 2005, 23(5): 612–616
https://doi.org/10.1038/nbt1083
14
Jantama K, Haupt M J, Svoronos S A, Zhang X L, Moore J C, Shanmugam K T, Ingram L O. Combining metabolic engineering and metabolic evolution to develop nonrecombinant strains of Escherichia coli C that produce succinate and malate. Biotechnology and Bioengineering, 2008, 99(5): 1140–1153
https://doi.org/10.1002/bit.21694
15
Dietrich J A, McKee A E, Keasling J D. High-throughput metabolic engineering: Advances in small-molecule screening and selection. Annual Review of Biochemistry, 2010, 79(1): 563–590
https://doi.org/10.1146/annurev-biochem-062608-095938
16
Kim Y, Ingram L O, Shanmugam K T. Construction of an Escherichia coli K-12 mutant for homoethanologenic fermentation of glucose or xylose without foreign genes. Applied and Environmental Microbiology, 2007, 73(6): 1766–1771
https://doi.org/10.1128/AEM.02456-06
17
Zhou S, Iverson A G, Grayburn W S. Engineering a native homoethanol pathway in Escherichia coli B for ethanol production. Biotechnology Letters, 2008, 30(2): 335–342
https://doi.org/10.1007/s10529-007-9544-x
18
Solem C, Dehli T, Jensen P R. Rewiring Lactococcus lactis for ethanol production. Applied and Environmental Microbiology, 2013, 79(8): 2512–2518
https://doi.org/10.1128/AEM.03623-12
19
Shen C R, Lan E I, Dekishima Y, Baez A, Cho K M, Liao J C. Driving forces enable high-titer anaerobic L-butanol synthesis in Escherichia coli. Applied and Environmental Microbiology, 2011, 77(9): 2905–2915
https://doi.org/10.1128/AEM.03034-10
20
Lim J H, Seo S W, Kim S Y, Jung G Y. Model-driven rebalancing of the intracellular redox state for optimization of a heterologous n-butanol pathway in Escherichia coli. Metabolic Engineering, 2013, 20: 49–55
https://doi.org/10.1016/j.ymben.2013.09.003
21
Yim H, Haselbeck R, Niu W, Pujol-Baxley C, Burgard A, Boldt J, Khandurina J, Trawick J D, Osterhout R E, Stephen R, . Metabolic engineering of Escherichia coli for direct production of 1,4-butanediol. Nature Chemical Biology, 2011, 7(7): 445–452
https://doi.org/10.1038/nchembio.580
22
Ida Y, Hirasawa T, Furusawa C, Shimizu H. Utilization of saccharomyces cerevisiae recombinant strain incapable of both ethanol and glycerol biosynthesis for anaerobic bioproduction. Applied Microbiology and Biotechnology, 2013, 97(11): 4811–4819
https://doi.org/10.1007/s00253-013-4760-x
23
Zhang X, Jantama K, Moore J C, Shanmugam K T, Ingram L O. Production of L-alanine by metabolically engineered Escherichia coli. Applied Microbiology and Biotechnology, 2007, 77(2): 355–366
https://doi.org/10.1007/s00253-007-1170-y
24
Jantama K, Zhang X, Moore J C, Shanmugam K T, Svoronos S A, Ingram L O. Eliminating side products and increasing succinate yields in engineered strains of Escherichia coli C. Biotechnology and Bioengineering, 2008, 101(5): 881–893
https://doi.org/10.1002/bit.22005
25
Klein-Marcuschamer D, Ajikumar P K, Stephanopoulos G. Engineering microbial cell factories for biosynthesis of isoprenoid molecules: Beyond lycopene. Trends in Biotechnology, 2007, 25(9): 417–424
https://doi.org/10.1016/j.tibtech.2007.07.006
26
Santos C N S, Stephanopoulos G. Melanin-based high-throughput screen for L-tyrosine production in Escherichia coli. Applied and Environmental Microbiology, 2008, 74(4): 1190–1197
https://doi.org/10.1128/AEM.02448-07
27
DeLoache W C, Russ Z N, Narcross L, Gonzales A M, Martin V J, Dueber J E. An enzyme-coupled biosensor enables (S)-reticuline production in yeast from glucose. Nature Chemical Biology, 2015, 11(7): 465–471
https://doi.org/10.1038/nchembio.1816
28
Binder S, Schendzielorz G, Stabler N, Krumbach K, Hoffmann K, Bott M, Eggeling L. A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level. Genome Biology, 2012, 13(5): 1
https://doi.org/10.1186/gb-2012-13-5-r40
29
Lin H, Tao H, Cornish V W. Directed evolution of a glycosynthase via chemical complementation. Journal of the American Chemical Society, 2004, 126(46): 15051–15059
https://doi.org/10.1021/ja046238v
30
Baker K, Bleczinski C, Lin H, Salazar-Jimenez G, Sengupta D, Krane S, Cornish V W. Chemical complementation: A reaction-independent genetic assay for enzyme catalysis. Proceedings of the National Academy of Sciences of the United States of America, 2002, 99(26): 16537–16542
https://doi.org/10.1073/pnas.262420099
31
Frommer W B, Davidson M W, Campbell R E. Genetically encoded biosensors based on engineered fluorescent proteins. Chemical Society Reviews, 2009, 38(10): 2833–2841
https://doi.org/10.1039/b907749a
32
Lalonde S, Ehrhardt D W, Frommer W B. Shining light on signaling and metabolic networks by genetically encoded biosensors. Current Opinion in Plant Biology, 2005, 8(6): 574–581
https://doi.org/10.1016/j.pbi.2005.09.015
33
Okumoto S, Looger L L, Micheva K D, Reimer R J, Smith S J, Frommer W B. Detection of glutamate release from neurons by genetically encoded surface-displayed FRET nanosensors. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(24): 8740–8745
https://doi.org/10.1073/pnas.0503274102
34
de Lorimier R M, Smith J J, Dwyer M A, Looger L L, Sali K M, Paavola C D, Rizk S S, Sadigov S, Conrad D W, Loew L, . Construction of a fluorescent biosensor family. Protein Science, 2002, 11(11): 2655–2675
https://doi.org/10.1110/ps.021860
35
Fehr M, Lalonde S, Lager I, Wolff M W, Frommer W B. In vivo imaging of the dynamics of glucose uptake in the cytosol of COS-7 cells by fluorescent nanosensors. Journal of Biological Chemistry, 2003, 278(21): 19127–19133
https://doi.org/10.1074/jbc.M301333200
36
Fehr M, Takanaga H, Ehrhardt D W, Frommer W B. Evidence for high-capacity bidirectional glucose transport across the endoplasmic reticulum membrane by genetically encoded fluorescence resonance energy transfer nanosensors. Molecular and Cellular Biology, 2005, 25(24): 11102–11112
https://doi.org/10.1128/MCB.25.24.11102-11112.2005
37
Kaper T, Lager I, Looger L L, Chermak D, Frommer W B. Fluorescence resonance energy transfer sensors for quantitative monitoring of pentose and disaccharide accumulation in bacteria. Biotechnology for Biofuels, 2008, 1(1): 1
https://doi.org/10.1186/1754-6834-1-11
38
Fehr M, Frommer W B, Lalonde S. Visualization of maltose uptake in living yeast cells by fluorescent nanosensors. Proceedings of the National Academy of Sciences of the United States of America, 2002, 99(15): 9846–9851
https://doi.org/10.1073/pnas.142089199
39
Deuschle K, Okumoto S, Fehr M, Looger L L, Kozhukh L, Frommer W B. Construction and optimization of a family of genetically encoded metabolite sensors by semirational protein engineering. Protein Science, 2005, 14(9): 2304–2314
https://doi.org/10.1110/ps.051508105
40
Okada S, Ota K, Ito T. Circular permutation of ligand-binding module improves dynamic range of genetically encoded FRET-based nanosensor. Protein Science, 2009, 18(12): 2518–2527
https://doi.org/10.1002/pro.266
Yang J, Seo S W, Jang S, Shin S I, Lim C H, Roh T Y, Jung G Y. Synthetic RNA devices to expedite the evolution of metabolite-producing microbes. Nature Communications, 2013, 4: 7
https://doi.org/10.1038/ncomms2404
43
Wachsmuth M, Findeiss S, Weissheimer N, Stadler P F, Morl M. De novo design of a synthetic riboswitch that regulates transcription termination. Nucleic Acids Research, 2013, 41(4): 2541–2551
https://doi.org/10.1093/nar/gks1330
44
Trausch J J, Ceres P, Reyes F E, Batey R T. The structure of a tetrahydrofolate-sensing riboswitch reveals two ligand binding sites in a single aptamer. Structure (London, England), 2011, 19(10): 1413–1423
https://doi.org/10.1016/j.str.2011.06.019
45
Desai S K, Gallivan J P. Genetic screens and selections for small molecules based on a synthetic riboswitch that activates protein translation. Journal of the American Chemical Society, 2004, 126(41): 13247–13254
https://doi.org/10.1021/ja048634j
46
Win M N, Smolke C D. Higher-order cellular information processing with synthetic RNA devices. Science, 2008, 322(5900): 456–460
https://doi.org/10.1126/science.1160311
47
Michener J K, Smolke C D. High-throughput enzyme evolution in Saccharomyces cerevisiae using a synthetic RNA switch. Metabolic Engineering, 2012, 14(4): 306–316
https://doi.org/10.1016/j.ymben.2012.04.004
48
Eckdahl T T, Campbell A M, Heyer L J, Poet J L, Blauch D N, Snyder N L, Atchley D T, Baker E J, Brown M, Brunner E C, . Programmed evolution for optimization of orthogonal metabolic output in bacteria. PLoS One, 2015, 10(2): 0118322
https://doi.org/10.1371/journal.pone.0118322
49
Ellington A D, Szostak J W. In vitro selection of RNA molecules that bind specific ligands. Nature, 1990, 346(6287): 818–822
https://doi.org/10.1038/346818a0
50
Win M N, Smolke C D. A modular and extensible RNA-based gene-regulatory platform for engineering cellular function. Proceedings of the National Academy of Sciences of the United States of America, 2007, 104(36): 14283–14288
https://doi.org/10.1073/pnas.0703961104
51
Ouellet J. RNA Fluorescence with light-up aptamers. Frontiers in Chemistry, 2016, 4: 29
52
Nakayama S, Luo Y, Zhou J, Dayie T K, Sintim H O. Nanomolar fluorescent detection of c-di-GMP using a modular aptamer strategy. Chemical Communications, 2012, 48(72): 9059–9061
https://doi.org/10.1039/c2cc34379g
53
Wang X C, Wilson S C, Hammond M C. Next-generation RNA-based fluorescent biosensors enable anaerobic detection of cyclic di-GMP. Nucleic Acids Research, 2016, 44(17): e139–e139
https://doi.org/10.1093/nar/gkw580
54
Kellenberger C A, Hammond M C. In vitro analysis of riboswitch-Spinach aptamer fusions as metabolite-sensing fluorescent biosensors. Methods in Enzymology, 2015, 550: 147–172
https://doi.org/10.1016/bs.mie.2014.10.045
55
Paige J S, Nguyen-Duc T, Song W, Jaffrey S R. Fluorescence imaging of cellular metabolites with RNA. Science, 2012, 335(6073): 1194
https://doi.org/10.1126/science.1218298
56
Su Y, Hickey S F, Keyser S G, Hammond M C. In vitro and in vivo enzyme activity screening via RNA-based fluorescent biosensors for S-adenosyl-L-homocysteine (SAH). Journal of the American Chemical Society, 2016, 138(22): 7040–7047
https://doi.org/10.1021/jacs.6b01621
57
Kellenberger C A, Chen C, Whiteley A T, Portnoy D A, Hammond M C. RNA-based fluorescent biosensors for live cell imaging of second messenger cyclic di-AMP. Journal of the American Chemical Society, 2015, 137(20): 6432–6435
https://doi.org/10.1021/jacs.5b00275
58
Binder S, Siedler S, Marienhagen J, Bott M, Eggeling L. Recombineering in corynebacterium glutamicum combined with optical nanosensors: A general strategy for fast producer strain generation. Nucleic Acids Research, 2013, 41(12): 6360–6369
https://doi.org/10.1093/nar/gkt312
59
Schendzielorz G, Dippong M, Grunberger A, Kohlheyer D, Yoshida A, Binder S, Nishiyama C, Nishiyama M, Bott M, Eggeling L. Taking control over control: Use of product sensing in single cells to remove flux control at key enzymes in biosynthesis pathways. ACS Synthetic Biology, 2014, 3(1): 21–29
https://doi.org/10.1021/sb400059y
60
Lange C, Mustafi N, Frunzke J, Kennerknecht N, Wessel M, Bott M, Wendisch V F. Lrp of Corynebacterium glutamicum controls expression of the brnFE operon encoding the export system for L-methionine and branched-chain amino acids. Journal of Biotechnology, 2012, 158(4): 231–241
https://doi.org/10.1016/j.jbiotec.2011.06.003
61
Mustafi N, Grunberger A, Kohlheyer D, Bott M, Frunzke J. The development and application of a single-cell biosensor for the detection of L-methionine and branched-chain amino acids. Metabolic Engineering, 2012, 14(4): 449–457
https://doi.org/10.1016/j.ymben.2012.02.002
62
Mahr R, Gatgens C, Gatgens J, Polen T, Kalinowski J, Frunzke J. Biosensor-driven adaptive laboratory evolution of L-valine production in Corynebacterium glutamicum. Metabolic Engineering, 2015, 32: 184–194
https://doi.org/10.1016/j.ymben.2015.09.017
63
Mustafi N, Grunberger A, Mahr R, Helfrich S, Noh K, Blombach B, Kohlheyer D, Frunzke J. Application of a genetically encoded biosensor for live cell imaging of L-valine production in pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum strains. PLoS One, 2014, 9(1): e85731
https://doi.org/10.1371/journal.pone.0085731
64
Bogner M, Ludewig U. Visualization of arginine influx into plant cells using a specific FRET-sensor. Journal of Fluorescence, 2007, 17(4): 350–360
https://doi.org/10.1007/s10895-007-0192-2
65
Mohsin M, Ahmad A. Genetically-encoded nanosensor for quantitative monitoring of methionine in bacterial and yeast cells. Biosensors & Bioelectronics, 2014, 59: 358–364
https://doi.org/10.1016/j.bios.2014.03.066
66
Mohsin M, Abdin M Z, Nischal L, Kardam H, Ahmad A. Genetically encoded FRET-based nanosensor for in vivo measurement of leucine. Biosensors & Bioelectronics, 2013, 50: 72–77
https://doi.org/10.1016/j.bios.2013.06.028
67
Wang J M, Gao D F, Yu X L, Li W, Qi Q S. Evolution of a chimeric aspartate kinase for L-lysine production using a synthetic RNA device. Applied Microbiology and Biotechnology, 2015, 99(20): 8527–8536
https://doi.org/10.1007/s00253-015-6615-0
68
Liu Y N, Li Q G, Zheng P, Zhang Z D, Liu Y F, Sun C M, Cao G Q, Zhou W J, Wang X W, Zhang D W, . Developing a high-throughput screening method for threonine overproduction based on an artificial promoter. Microbial Cell Factories, 2015, 14(1): 1
https://doi.org/10.1186/s12934-015-0311-8
69
Zaslaver A, Bren A, Ronen M, Itzkovitz S, Kikoin I, Shavit S, Liebermeister W, Surette M G, Alon U. A comprehensive library of fluorescent transcriptional reporters for Escherichia coli. Nature Methods, 2006, 3(8): 623–628
https://doi.org/10.1038/nmeth895
70
Mahr R, von Boeselager R F, Wiechert J, Frunzke J. Screening of an Escherichia coli promoter library for a phenylalanine biosensor. Applied Microbiology and Biotechnology, 2016, 100(15):6739–6753
71
Dietrich J A, Shis D L, Alikhani A, Keasling J D. Transcription factor-based screens and synthetic selections for microbial small-molecule biosynthesis. ACS Synthetic Biology, 2013, 2(1): 47–58
https://doi.org/10.1021/sb300091d
72
Szmidt-Middleton H L, Ouellet M, Adams P D, Keasling J D, Mukhopadhyay A. Utilizing a highly responsive gene, yhjX, in E. coli based production of 1,4-butanediol. Chemical Engineering Science, 2013, 103: 68–73
https://doi.org/10.1016/j.ces.2013.06.044
73
Uchiyama T, Miyazaki K. Product-induced gene expression, a product-responsive reporter assay used to screen metagenomic libraries for enzyme-encoding genes. Applied and Environmental Microbiology, 2010, 76(21): 7029–7035
https://doi.org/10.1128/AEM.00464-10
74
van Sint Fiet S, van Beilen J B, Witholt B. Selection of biocatalysts for chemical synthesis. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(6): 1693–1698
https://doi.org/10.1073/pnas.0504733102
75
Raman S, Rogers J K, Taylor N D, Church G M. Evolution-guided optimization of biosynthetic pathways. Proceedings of the National Academy of Sciences of the United States of America, 2014, 111(50): 17803–17808
https://doi.org/10.1073/pnas.1409523111
76
Chen W, Zhang S, Jiang P X, Yao J, He Y Z, Chen L C, Gui X W, Dong Z Y, Tang S Y. Design of an ectoine-responsive AraC mutant and its application in metabolic engineering of ectoine biosynthesis. Metabolic Engineering, 2015, 30: 149–155
https://doi.org/10.1016/j.ymben.2015.05.004
77
Mukherjee K, Bhattacharyya S, Peralta-Yahya P. GPCR-based chemical biosensors for medium-chain fatty acids. ACS Synthetic Biology, 2015, 4(12): 1261–1269
https://doi.org/10.1021/sb500365m
78
Tang S Y, Cirino P C. Design and application of a mevalonate-responsive regulatory protein. Angewandte Chemie International Edition, 2011, 50(5): 1084–1086
https://doi.org/10.1002/anie.201006083
79
Tang S Y, Qian S, Akinterinwa O, Frei C S, Gredell J A, Cirino P C. Screening for enhanced triacetic acid lactone production by recombinant Escherichia coli expressing a designed triacetic acid lactone reporter. Journal of the American Chemical Society, 2013, 135(27): 10099–10103
https://doi.org/10.1021/ja402654z
80
Hichri I, Barrieu F, Bogs J, Kappel C, Delrot S, Lauvergeat V. Recent advances in the transcriptional regulation of the flavonoid biosynthetic pathway. Journal of Experimental Botany, 2011, 62(8): 2465–2483
https://doi.org/10.1093/jxb/erq442
81
Siedler S, Stahlhut S G, Malla S, Maury J, Neves A R. Novel biosensors based on flavonoid-responsive transcriptional regulators introduced into Escherichia coli. Metabolic Engineering, 2014, 21: 2–8
https://doi.org/10.1016/j.ymben.2013.10.011
82
Marin A M, Souza E M, Pedrosa F O, Souza L M, Sassaki G L, Baura V A, Yates M G, Wassem R, Monteiro R A. Naringenin degradation by the endophytic diazotroph Herbaspirillum seropedicae SmR1. Microbiology, 2013, 159(1): 167–175
https://doi.org/10.1099/mic.0.061135-0
83
Teran W, Felipe A, Segura A, Rojas A, Ramos J L, Gallegos M T. Antibiotic-dependent induction of Pseudomonas putida DOT-T1E TtgABC efflux pump is mediated by the drug binding repressor TtgR. Antimicrobial Agents and Chemotherapy, 2003, 47(10): 3067–3072
https://doi.org/10.1128/AAC.47.10.3067-3072.2003
84
Jenison R D, Gill S C, Pardi A, Polisky B. High-resolution molecular discrimination by RNA. Science, 1994, 263(5152): 1425–1429
https://doi.org/10.1126/science.7510417
85
Thompson K M, Syrett H A, Knudsen S M, Ellington A D. Group I aptazymes as genetic regulatory switches. BMC Biotechnology, 2002, 2(1): 1
https://doi.org/10.1186/1472-6750-2-21
86
Chou H H, Keasling J D. Programming adaptive control to evolve increased metabolite production. Nature Communications, 2013, 4: 8
https://doi.org/10.1038/ncomms3595
87
Park Y H, Koo H M, Moon J O, Kim S J, Kim H J, Lee J K. L-Lysine-inducible promoter. US 07851198, <Date>Dec 14 2010</Date>, 2010
88
Wang Y, Li Q, Zheng P, Guo Y, Wang L, Zhang T, Sun J, Ma Y. Evolving the L-lysine high-producing strain of Escherichia coli using a newly developed high-throughput screening method. Journal of Industrial Microbiology & Biotechnology, 2016, 43(9): 1227–1235
https://doi.org/10.1007/s10295-016-1803-1
89
Kim Y S, Gu M B. Advances in aptamer screening and small molecule aptasensors. Biosensors Based on Aptamers and Enzymes, 2014, 140: 29–67
https://doi.org/10.1007/10_2013_225
90
Ruscito A, DeRosa M C. Small-molecule binding aptamers: Selection strategies, characterization, and applications. Frontiers in Chemistry, 2016, 4: 14
91
McKeague M, Derosa M C. Challenges and opportunities for small molecule aptamer development. Journal of Nucleic Acids, 2012, 2012: 748913