eISSN: 2221-6197 DOI: 10.31301/2221-6197

Natural and synthetic indigoids as compounds of a wide range of applications

Year: 2025

Pages: 263-274

Number: Volume 17, issue 3

Type: scientific article

Summary:

It is now known that the blue pigment indigo (in particular, its precursors) is found in about 300 plant species, but only a few species serve as the main sources of indigo, including Indigofera tinctorium, Isatis tinctorium, Polygonum tinctorium (Persikaria tinctoria), Strobilanthes cusia (Baphicacanthus cusia). For several millennia, mankind has been using the plant pigment indigo to dye fabrics and color human bodies. With the development of civilization, indigo also began to be used in traditional medicine to treat various diseases. For many years, indigo was extracted from plant raw materials, but at the end of the 19th century, chemical synthesis of indigo was developed. Nowadays, natural indigo has been almost completely replaced by industrially produced one. About 95% of all synthesized dye is spent on dyeing textile materials, primarily denim, which is used for sewing jeans. The water–soluble indigo derivative, indigocarmine, is employed in medicine, as well as in the food industry as a food additive E132 to give products a cyan or blue color. In the new millennium, due to advances in biochemistry and molecular biology, which revealed the pathways of indigoid biosynthesis in plants and cloned genes of enzymes involved in indigo metabolism, it became possible to use indigo as a reporter molecule in the creation of transgenic plants. The electronics industry also began to show interest in indigo.

Keywords:

indigo, indigoids, indigotin, indican, indoxyl, flavin-dependent monooxygenase, transgenic plants, reporter genes

References:

  1. Abad-Valle P, Fernández-Abedul MT, Costa-García A. Genosensor on gold films with enzymatic electrochemical detection of a SARS virus sequence. Biosens Bioelectron. 20(11). 2251-2260. doi: 10.1016/j.bios.2004.10.019
  2. Abad-Valle P, Fernández-Abedul MT, Costa-García A. DNA single-base mismatch study with an electrochemical enzymatic genosensor. Biosens Bioelectron. 22(8). 1642-1650. doi: 10.1016/j.bios.2006.07.015
  3. Albayrak İ, Cessur A, Demirci T et al. Enhancement of indigotin and indirubin production in root cultures of Isatis species by H2O2: biochemical and molecular responses. Sci Rep. 15(1). 33637. doi: 10.1038/s41598-025-10242-4
  4. Ashizawa M, Masuda N, Higashino T et al. Ambipolar organic transistors based on isoindigo derivatives. Organic Electronics. 2016. 35. 95-100. doi: 10.1016/j.orgel.2016.05.013
  5. Baymiev AlKh, Chemeris DA, Sakhabutdinova AR et al. In higher plants as an example, one can see that the era of sequencing of their diploid genomes is coming. Biomics. 2025. V.17(1). P. 17 – 41. DOI:31301/2221-6197.bmcs.2025-3
  6. Catucci G, Turella S, Cheropkina H et al. Green production of indigo and indirubin by an engineered Baeyer–Villiger monooxygenase. Biocatalysis and Agricultural Biotechnology. 44. 102458. doi: 10.1016/j.bcab.2022.102458
  7. Chandel N, Singh BB, Dureja C et al. Indigo production goes green: a review on opportunities and challenges of fermentative production. World J Microbiol Biotechnol. 40(2). 62. doi: 10.1007/s11274-023-03871-2
  8. Chen J, Dong X, Li Q et al. Biosynthesis of the active compounds of Isatis indigotica based on transcriptome sequencing and metabolites profiling. BMC Genomics. 14. 857. doi: 10.1186/1471-2164-14-857
  9. Chen T, Wang X, Zhuang L et al. Development and optimization of a microbial co-culture system for heterologous indigo biosynthesis. Microb Cell Fact. 20(1). 154. doi: 10.1186/s12934-021-01636-w
  10. Choi K-Y. A review of recent progress in the synthesis of bio-indigoids and their biologically assisted end-use applications. Dyes and Pigments. 181. 108570. doi: 10.1016/j.dyepig.2020.108570
  11. Choi HS, Kim JK, Cho EH et al. A novel flavin-containing monooxygenase from Methylophaga sp strain SK1 and its indigo synthesis in Escherichia coli. Biochem Biophys Res Commun. 2003. 306(4). 930-936. doi: 10.1016/s0006-291x(03)01087-8
  12. Cooksey CJ. An annotated bibliography of recent significant publications on indigo and related compounds. Biotech Histochem. 87(7). 439-463. doi: 10.3109/10520295.2012.698308
  13. Devi RL, Devi ST, Sanatombi K. Hairy root culture of Strobilanthes cusia for the production and enhancement of indigo biosynthesis. Plant Cell, Tissue and Organ Culture. 157. 62. doi: 10.1007/s11240-024-02791-9
  14. Díaz-González M, de la Escosura-Muñiz A, González-García MB et al. DNA hybridization biosensors using polylysine modified SPCEs. Biosens Bioelectron. 23(9). 1340-1346. doi: 10.1016/j.bios.2007.12.001
  15. Fan C, Xie Z, Zheng D et al. Overview of indigo biosynthesis by Flavin-containing Monooxygenases: History, industrialization challenges, and strategies. Biotechnol Adv. 73. 108374. doi: 10.1016/j.biotechadv.2024.108374
  16. Głowacki ED, Voss G, Demirak K et al. A facile protection–deprotection route for obtaining indigo pigments as thin films and their applications in organic bulk heterojunctions. Commun. 2013a. 49(54). 6063-6065. doi: 10.1039/C3CC42889C
  17. Głowacki ED, Voss G, Sariciftci NS. 25th anniversary article: progress in chemistry and applications of functional indigos for organic electronics. Adv Mater. 25(47). 6783-6800. doi: 10.1002/adma.201302652
  18. Gray PMM. The formation of indigotin from indole by soil bacteria. Proc Roy Soc Lond Ser B. 1928. 102. 263–280. doi: 10.1098/rspb.1928.0003
  19. Hartl A, Polleichtner A, Novak J. "Purplish Blue" or "Greenish Grey"? Indigo Qualities and Extraction Yields from Six Species. Plants (Basel). 13(7). 918. doi: 10.3390/plants13070918
  20. Heo B-G, Park Y-J, Park Y-S et al. Anticancer and antioxidant effects of extracts from different parts of indigo plant. Industrial Crops and Products. 2014. 56. 9-16. doi: 10.1016/j.indcrop.2014.02.023
  21. Hsu TM, Welner DH, Russ ZN et al. Employing a biochemical protecting group for a sustainable indigo dyeing strategy. Nat Chem Biol. 14(3). 256-261. doi: 10.1038/nchembio.2552
  22. Inoue S, Morita R, Kuwata K et al. Tissue-specific and intracellular localization of indican synthase from Polygonum tinctorium. Plant Physiol Biochem. 132. 138-144. doi: 10.1016/j.plaphy.2018.08.034
  23. Inoue S, Morita R, Minami Y. An indigo-producing plant, Polygonum tinctorium, possesses a flavin-containing monooxygenase capable of oxidizing indole. Biochem Biophys Res Commun. 2021. 534. 199-205. doi: 10.1016/j.bbrc.2020.11.112
  24. Inoue S, Moriya T, Morita R et al. Characterization of UDP-glucosyltransferase from Indigofera tinctoria. Plant Physiol Biochem. 121. 226-233. doi: 10.1016/j.plaphy.2017.11.002
  25. Irimia-Vladu M, Głowacki ED, Troshin PA et al. Indigo--a natural pigment for high performance ambipolar organic field effect transistors and circuits. Adv Mater. 24(3). 375-80. doi: 10.1002/adma.201102619
  26. Jiao J, Gai QY, Yao LP et al. Ultraviolet radiation for flavonoid augmentation in Isatis tinctoria L. hairy root cultures mediated by oxidative stress and biosynthetic gene expression. Ind Crops Prod. 118. 347-354. doi: 10.1016/j.indcrop.2018.03.046
  27. Kaplan G, Seferoğlu Z, Berdnikova DV. Photochromic derivatives of indigo: historical overview of development, challenges and applications. Beilstein J Org Chem. 20. 228-242. doi: 10.3762/bjoc.20.23
  28. Khan MI, Polturak G. Biotechnological production and emerging applications of betalains: A review. Biotechnol Adv. 81. 108576. doi: 10.1016/j.biotechadv.2025.108576
  29. Kim HJ, Lee Y, Shin Y et al. Finding of the positive impact of glucose on the production of indican over indigo in engineered Escherichia coli. J Ind Microbiol Biotechnol. 52. kuae048. doi: 10.1093/jimb/kuae048
  30. Langer P. N-Glycosides of indigo, indirubin, and isoindigo: blue, red, and yellow sugars and their cancerostatic activity. Beilstein J Org Chem. 20. 2840-2869. doi: 10.3762/bjoc.20.240
  31. Larkin A, Harbison S. An improved method for STR analysis of bloodstained denim. Int J Legal Med. 112(6). 388-390. doi: 10.1007/s004140050020
  32. Li Q, Huang H, Fan R et al. Chromosome-level genome assembly of the tetraploid medicinal and natural dye plant Persicaria tinctoria. Sci Data. 2024. 11(1). 1440. doi: 10.1038/s41597-024-04317-6
  33. Li S, Cunningham AB, Fan R et al. Identity blues: the ethnobotany of the indigo dyeing by Landian Yao (Iu Mien) in Yunnan, Southwest China. J Ethnobiol Ethnomed. 15(1). 13. doi: 10.1186/s13002-019-0289-0
  34. Lin W, Huang W, Ning S et al. De novo characterization of the Baphicacanthus cusia (Nees) Bremek transcriptome and analysis of candidate genes involved in indican biosynthesis and metabolism. PLoS One. 13(7). e0199788. doi: 10.1371/journal.pone.0199788
  35. Linke JA, Rayat A, Ward JM. Production of indigo by recombinant bacteria. Bioresour Bioprocess. 2023. 10(1). 20. doi: 10.1186/s40643-023-00626-7
  36. Lopes HFS, Tu Z, Sumi H e al. Indigofera tinctoria L. leaf powder promotes initiation of indigo reduction by inducing of rapid transition of the microbial community. Front Microbiol. 13. 957809. doi: 10.3389/fmicb.2022.957809
  37. Ma Q, Qu Y, Tang H et al. Genome sequence of a novel indigo-producing strain, Pseudomonas monteilii QM. J Bacteriol. 2012. 194(16). 4459-4460. doi: 10.1128/JB.00867-12
  38. Ma Y, Zhang G, Pan J. Spectroscopic studies of DNA interactions with food colorant indigo carmine with the use of ethidium bromide as a fluorescence probe. J Agric Food Chem. 2012. 60(43). 10867-10875. doi: 10.1021/jf303698k
  39. Manivannan R, Prabakaran K, SonY-A. Eco-friendly approach of bio-indigo synthesis and developing purification methods towards isolation of indigo from indirubin and bacterial fragments. Sustainable Chemistry and Pharmacy. 2025. 44(3). 101954. doi: 10.1016/j.scp.2025.101954
  40. Minami Y, Nishimura O, Hara-Nishimura I et al. Tissue and intracellular localization of indican and the purification and characterization of indican synthase from indigo plants. Plant Cell Physiol. 2000. 41(2). 218-25. doi: 10.1093/pcp/41.2.218
  41. Minami Y, Shigeta Y, Tokumoto U et al. Cloning, sequencing, characterization, and expression of a β-glucosidase cDNA from the indigo plant. Plant Science. 142(2). 219-226. doi: 10.1016/S0168-9452(99)00015-1.
  42. Minami Y, Takao H, Kanafuji T et al. beta-Glucosidase in the indigo plant: intracellular localization and tissue specific expression in leaves. Plant Cell Physiol. 38(9). 1069-1074. doi: 10.1093/oxfordjournals.pcp.a029273
  43. O'Connor KE, Dobson AD, Hartmans S. Indigo formation by microorganisms expressing styrene monooxygenase activity. Appl Environ Microbiol. 1997. 63(11). 4287-4291. doi: 10.1128/aem.63.11.4287-4291.1997
  44. Patolsky F, Lichtenstein A, Willner I. Highly sensitive amplified electronic detection of DNA by biocatalyzed precipitation of an insoluble product onto electrodes. Chemistry. 2003. 9(5). 1137-1145. doi: 10.1002/chem.200390131
  45. Pham NN, Wu YH, Dai TA et al. Auto-inducible synthetic pathway in coli enhanced sustainable indigo production from glucose. Metab Eng. 2024. 85. 14-25. doi: 10.1016/j.ymben.2024.07.002
  46. Qi-Yue Y, Ting Z, Ya-Nan H et al. From natural dye to herbal medicine: a systematic review of chemical constituents, pharmacological effects and clinical applications of indigo naturalis. Chin Med. 15(1). 127. doi: 10.1186/s13020-020-00406-x
  47. Qu Y, Ma Q, Zhang X et al. Optimization of indigo production by a newly isolated Pseudomonas sp. J Basic Microbiol. 2012. 52(6). 687-694. doi: 10.1002/jobm.201100516
  48. Qu Y, Pi W, Ma F et al. Influence and optimization of growth substrates on indigo formation by a novel isolate Acinetobacter sp. PP-2. Bioresour Technol. 2010. 101(12). 4527-4532. doi: 10.1016/j.biortech.2010.01.033
  49. Qu Y, Zhang X, Ma Q et al. Indigo biosynthesis by Comamonas sp. Biotechnol Lett. 2012. 34(2). 353-357. doi: 10.1007/s10529-011-0778-2
  50. Rajan AK, Cindrella L. Studies on new natural dye sensitizers from Indigofera tinctoria in dye-sensitized solar cells. Optical Materials. 2019. 88. 39-47. doi: 10.1016/j.optmat.2018.11.016
  51. Raza S, Bończak B, Atamas N et al. The activity of indigo carmine against bacteriophages: an edible antiphage agent. Appl Microbiol Biotechnol. 109(1). 24. doi: 10.1007/s00253-025-13414-4
  52. Sagwan-Barkdoll L, Kim MJ, Berim A et al. Plant P450 forms indigo and indirubin when expressed in Escherichia coli. 2025. 229. 114268. doi: 10.1016/j.phytochem.2024.114268
  53. Sarangi BK, Minami Y, Thul ST. RNA-Seq analysis for indigo biosynthesis pathway genes in Indigofera tinctoria and Polygonum tinctorium. Genom Data. 2015. 6. 212-3. doi: 10.1016/j.gdata.2015.09.021
  54. Splitstoser JC, Dillehay TD, Wouters J et al. Early pre-Hispanic use of indigo blue in Peru. Sci Adv. 2016. 2(9). e1501623. doi: 10.1126/sciadv.1501623
  55. Stasiak N, Kukuła-Koch W, Głowniak K. Modern industrial and pharmacological applications of indigo dye and its derivatives--a review. Acta Pol Pharm. 2014. 71(2). 215-221.
  56. Súnico V, Piunti I, Bhattacharjee M et al. Overview on Current Selectable Marker Systems and Novel Marker Free Approaches in Fruit Tree Genetic Engineering. Int J Mol Sci. 25(22). 11902. doi: 10.3390/ijms252211902
  57. Tang X, Xiao Y, Lv T et al. High-throughput sequencing and De Novo assembly of the Isatis indigotica PLoS One. 2014. 9(9). :e102963. doi: 10.1371/journal.pone.0102963
  58. Wang P, Si H, Li C et al. Plant genetic transformation: achievements, current status and future prospects. Plant Biotechnol J. 23(6). 2034-2058. doi: 10.1111/pbi.70028
  59. Wang Z, Sierros K, Seehra MS et al. Development of indigo-based nonvolatile write-once-read-many-times memory device. Materials Letters. 2017. 206. 128-131. doi: 10.1016/j.matlet.2017.06.122
  60. Willner I, Patolsky F, Weizmann Y et al. Amplified detection of single-base mismatches in DNA using microgravimetric quartz-crystal-microbalance transduction. Talanta. 2002. 56(5). 847-856. doi: 10.1016/s0039-9140(01)00658-0
  61. Witty M. Thaumatin II: a simple marker gene for use in plants. Nucleic Acids Res. 1989. 17(8). 3312. doi: 10.1093/nar/17.8.3312
  62. Witty M. Thaumatin II: a sweet marker gene for use in plants. Methods Enzymol. 1992. 216. 441-447. doi: 10.1016/0076-6879(92)16040-q
  63. Xu W, Zhang L, Cunningham AB et al. Blue genome: chromosome-scale genome reveals the evolutionary and molecular basis of indigo biosynthesis in Strobilanthes cusia. Plant J. 2020. 104(4). 864-879. doi: 10.1111/tpj.14992
  64. Yin H, Chen H, Yan M et al. Efficient Bioproduction of Indigo and Indirubin by Optimizing a Novel Terpenoid Cyclase XiaI in Escherichia coli. ACS Omega. 2021. 6(31). 20569-20576. doi: 10.1021/acsomega.1c02679
  65. Yao M, Araki M, Senoh H et al. Indigo Dye as a Positive-electrode Material for Rechargeable Lithium Batteries. Chemistry Letters. 2010. 39(9). 950–952. doi: 10.1246/cl.2010.950
  66. Yu J, Zhang Y, Ning S et al. Molecular cloning and metabolomic characterization of the 5-enolpyruvylshikimate-3-phosphate synthase gene from Baphicacanthus cusia. BMC Plant Biol. 19(1). 485. doi: 10.1186/s12870-019-2035-0
  67. Zawirska-Wojtasiak R, Gośliński M, Szwacka M et al. Aroma evaluation of transgenic, thaumatin II-producing cucumber fruits. J Food Sci. 2009. 74(3). C204-210. doi: 10.1111/j.1750-3841.2009.01082.x
  68. Zhang YM, Su Y, Dai ZW et al. Integration of the metabolome and transcriptome reveals indigo biosynthesis in Phaius flavus flowers under freezing treatment. PeerJ. 2022. 10. e13106. doi: 10.7717/peerj.13106
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eISSN: 2221-6197 DOI: 10.31301/2221-6197