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Recent metabolomics and gene editing approaches for synthesis of microbial secondary metabolites for drug discovery and development

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Abstract

Microbial secondary metabolites (SMs) have been identified as an important natural source of drugs for several metabolic and neurological diseases. Along with biomedical applications, SMs are also used in the food and biochemical industries. SMs include natural products such as pigments, alkaloids, toxins, antimicrobials obtained from cultured microorganisms, while other non-cultivable microorganisms have also acted as a rich source of SMs. But, the isolation of SMs from these sources is a very tedious task. Metabolomics provides complete identification and structural information about the entire cellular metabolome under specific conditions using highly sophisticated instrumentation. Further, gene editing techniques such as cloning and gene refactoring, including advanced CRISPR-Cas, can be used for engineering microbes that have the potential to produce natural SMs that were not produced in native microbial strain. The present review describes integrated metabolomics and gene editing approaches for the synthesis of novel microbial SMs and their potential application towards drug discovery and development.

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References

  • Alvarez-Rivera G, Ballesteros-Vivas D, Parada-Alfonso F, Ibañez E, Cifuentes A (2019) Recent applications of high resolution mass spectrometry for the characterization of plant natural products. Trends Anal Chem 112:87–101

    CAS  Google Scholar 

  • Antón J, Lucio M, Peña A, Cifuentes A, Brito-Echeverría J, Moritz F, Tziotis D, López C, Urdiain M, Schmitt-Kopplin P, Rosselló-Móra R (2013) High metabolomic microdiversity within co-occurring isolates of the extremely halophilic bacterium Salinibacter ruber. PLoS ONE 8(5):e64701

    PubMed  PubMed Central  Google Scholar 

  • Baidoo EEK (2019) Microbial metabolomics: a general overview. Methods Mol Biol 1859:1–8

    CAS  PubMed  Google Scholar 

  • Bauman KD, Li J, Murata K, Mantovani SM, Dahesh S, Nizet V, Luhavaya H, Moore BS (2019) Refactoring the cryptic streptophenazine biosynthetic gene cluster unites phenazine, polyketide, and nonribosomal peptide biochemistry. Cell Chem Biol 26:724–736

    CAS  PubMed  PubMed Central  Google Scholar 

  • Blaženović I, Kind T, Ji J, Fiehn O (2018) Software tools and approaches for compound identification of LC-MS/MS data in metabolomics. Metabolites 8(2):31

    PubMed Central  Google Scholar 

  • Blin K, Kim HU, Medema MH, Weber T (2017) Recent development of antiSMASH and other computational approaches to mine secondary metabolite biosynthetic gene clusters. Brief Bioinform. https://doi.org/10.1093/bib/bbx146

    Article  PubMed Central  Google Scholar 

  • Bonet B, Teufel R, Crüsemann M, Ziemert N, Moore BS (2015) Direct capture and heterologous expression of Salinispora natural product genes for the biosynthesis of enterocin. J Nat Prod 78:539–542

    CAS  PubMed  Google Scholar 

  • Bose U, Hewavitharana A, Ng Y, Shaw P, Fuerst J, Hodson M (2015) LC-MS-based metabolomics study of marine bacterial secondary metabolite and antibiotic production in Salinispora arenicola. Mar Drugs 13:249–266

    PubMed  PubMed Central  Google Scholar 

  • Braaksma M, Bijlsma S, Coulier L, Punt PJ, van der Werf MJ (2011) Metabolomics as a tool for target identification in strain improvement: the influence of phenotype definition. Microbiology 157:147–159

    CAS  PubMed  Google Scholar 

  • Breitling R, Ceniceros A, Jankevics A, Takano E (2013) Metabolomics for secondary metabolite research. Metabolites 3:1076–1083

    CAS  PubMed  PubMed Central  Google Scholar 

  • Chaudhary T, Shukla P (2019) Bioinoculant capability enhancement through metabolomics and systems biology approaches. Brief Funct Genomics 18:159–168

    Google Scholar 

  • Cheng C, MacIntyre L, Abdelmohsen UR, Horn H, Polymenakou PN, Edrada-Ebel R, Hentschel U (2015) Biodiversity, anti-trypanosomal activity screening, and metabolomic profiling of actinomycetes isolated from Mediterranean sponges. PLoS ONE 10:0138528

    Google Scholar 

  • Chong J, Xia J (2018) MetaboAnalystR: an R package for flexible and reproducible analysis of metabolomics data. Bioinformatics 34:4313–4314

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cobb RE, Wang Y, Zhao H (2015) High-efficiency multiplex genome editing of Streptomyces species using an engineered CRISPR/Cas system. ACS Synth Biol 4:723–728

    CAS  PubMed  Google Scholar 

  • Cohen DR, Townsend CA (2018) A dual role for a polyketide synthase in dynemicinenediyne and anthraquinone biosynthesis. Nat Chem 10:231

    CAS  PubMed  Google Scholar 

  • Cortina NS, Krug D, Plaza A, Revermann O, Müller R (2012) Myxoprincomide: a natural product from Myxococcus xanthus discovered by comprehensive analysis of the secondary metabolome. Angew Chem 51:811–816

    CAS  Google Scholar 

  • Covington BC, McLean JA, Bachmann BO (2017) Comparative mass spectrometry-based metabolomics strategies for the investigation of microbial secondary metabolites. Nat Prod Rep 34:6–24

    CAS  PubMed  PubMed Central  Google Scholar 

  • D'Agostino P, Gulder TA (2018) Direct Pathway Cloning (DiPaC) combined with Sequence-and Ligation-Independent Cloning (SLIC) for fast biosynthetic gene cluster refactoring and heterologous expression. ACS Synth Biol 7:1702–1708

    CAS  PubMed  Google Scholar 

  • Dangi AK, Dubey KK, Shukla P (2017) Strategies to improve Saccharomyces cerevisiae: technological advancements and evolutionary engineering. Indian J Microbiol 57:378–386

    CAS  PubMed  PubMed Central  Google Scholar 

  • Dangi AK, Sinha R, Dwivedi S, Gupta SK, Shukla PS (2018) Cell line techniques and gene editing tools for antibody production: a review. Front Pharmacol 9:630

    PubMed  PubMed Central  Google Scholar 

  • Dhakal D, Dhakal Y, Sapkota D, Sohng JK (2017) Drug discovery based on microbial metabolism and communities. EC Microbiol (SI) 1:6–9

  • Dwivedi GR, Sisodia BS (2019) Secondary metabolites: metabolomics for secondary metabolites. In: Gupta VK, Pandey A (eds) New and future developments in microbial biotechnology and bioengineering. Elsevier, Amsterdam, pp 333–344

    Google Scholar 

  • Feling RH, Buchanan GO, Mincer TJ, Kauffman CA, Jensen PR, Fenical W (2003) Salinosporamide A: a highly cytotoxic proteasome inhibitor from a novel microbial source, a marine bacterium of the new genus Salinospora. Angew Chem 42:355–357

    CAS  Google Scholar 

  • Fillet M, Frédérich M (2015) The emergence of metabolomics as a key discipline in the drug discovery process. Drug Discov Today Technol 13:19–24

    PubMed  Google Scholar 

  • Greunke C, Duell ER, D’Agostino PM, Glöckle A, Lamm K, Gulder TAM (2018) Direct Pathway Cloning (DiPaC) to unlock natural product biosynthetic potential. Metab Eng 47:334–345

    CAS  PubMed  Google Scholar 

  • Harvey AL, Edrada-Ebel R, Quinn RJ (2015) The re-emergence of natural products for drug discovery in the genomics era. Nat Rev Drug Discov 14:111

    CAS  PubMed  Google Scholar 

  • Hollywood KA, Schmidt K, Takano E, Breitling R (2018) Metabolomics tools for the synthetic biology of natural products. Curr Opin Biotechnol 54:114–120

    CAS  PubMed  Google Scholar 

  • Horbal L, Marques F, Nadmid S, Mendes MV, Luzhetskyy A (2018) Secondary metabolites overproduction through transcriptional gene cluster refactoring. Metab Eng 49:299–315

    CAS  PubMed  Google Scholar 

  • Huang H, Zheng G, Jiang W, Hu H, Lu Y (2015) One-step high-efficiency CRISPR/Cas9-mediated genome editing in Streptomyces. Acta Biochim Biophys Sin 47:231–243

    CAS  PubMed  Google Scholar 

  • Jaddoa HH, Hameed IH, Mohammed GJ (2016) Analysis of volatile metabolites released by Staphylococcus aureus using gas chromatography-mass spectrometry and determination of its antifungal activity. Orient J Chem 32:2107–2116

    CAS  Google Scholar 

  • Jaiswal S, Singh DK, Shukla P (2019) Gene editing and systems biology tools for pesticide bioremediation: a review. Front Microbiol 10:87

    PubMed  PubMed Central  Google Scholar 

  • Jia H, Zhang L, Wang T, Han J, Tang H, Zhang L (2017) Development of a CRISPR/Cas9-mediated gene-editing tool in Streptomyces rimosus. Microbiology 163:1148–1155

    CAS  PubMed  Google Scholar 

  • Jiang W, Zhao X, Gabrieli T, Lou C, Ebenstein Y, Zhu TF (2015) Cas9-Assisted targeting of chromosome segments CATCH enables one-step targeted cloning of large gene clusters. Nat Commun 6:8101

    PubMed  Google Scholar 

  • Johnston CW, Connaty AD, Skinnider MA, Li Y, Grunwald A, Wyatt MA, Kerr RG, Magarvey NA (2016) Informatic search strategies to discover analogues and variants of natural product archetypes. J Ind Microbiol Biotechnol 43:293–298

    CAS  PubMed  Google Scholar 

  • Kang HS, Charlop-Powers Z, Brady SF (2016) Multiplexed CRISPR/Cas9-and TAR-mediated promoter engineering of natural product biosynthetic gene clusters in yeast. ACS Synth Biol 5:1002–1010

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kildgaard S, Mansson M, Dosen I, Klitgaard A, Frisvad JC, Larsen TO, Nielsen KF (2014) Accurate dereplication of bioactive secondary metabolites from marine-derived fungi by UHPLC-DAD-QTOFMS and a MS/HRMS library. Mar Drugs 12:3681–3705

    PubMed  PubMed Central  Google Scholar 

  • Krug D, Zurek G, Revermann O, Vos M, Velicer GJ, Müller R (2008) Discovering the hidden secondary metabolome of Myxococcus xanthus: a study of intraspecific diversity. Appl Environ Microbiol 74:3058–3068

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lee NC, Larionov V, Kouprina N (2015) Highly efficient CRISPR/Cas9-mediated TAR cloning of genes and chromosomal loci from complex genomes in yeast. Nucleic Acids Res 43:e55–e55

    PubMed  PubMed Central  Google Scholar 

  • Lee N, Hwang S, Lee Y, Cho S, Palsson B, Cho BK (2019a) Synthetic biology tools for novel secondary metabolite discovery in Streptomyces. J Microbiol Biotechnol 29:667–686

    PubMed  Google Scholar 

  • Lee SY, Mediani A, Ismail IS, Abas F (2019b) Antioxidants and α-glucosidase inhibitors from Neptunia oleracea fractions using 1 H NMR-based metabolomics approach and UHPLC-MS/MS analysis. BMC Complement Altern Med 19:7

    PubMed  PubMed Central  Google Scholar 

  • Lenders J, Frédérich M, De Tullio P (2015) Nuclear magnetic resonance: a key metabolomics platform in the drug discovery process. Drug Discov Today Technol 13:39–46

    Google Scholar 

  • Li L, Zheng G, Chen J, Ge M, Jiang W, Lu Y (2017) Multiplexed site-specific genome engineering for overproducing bioactive secondary metabolites in actinomycetes. Metab Eng 40:80–92

    CAS  PubMed  Google Scholar 

  • Liu Y, Tao W, Wen S, Li Z, Yang A, Deng Z, Sun Y (2015) In vitro CRISPR/Cas9 system for efficient targeted DNA editing. Mbio 6:e01714–e1715

    CAS  PubMed  PubMed Central  Google Scholar 

  • Low ZJ, Pang LM, Ding Y, Cheang QW, Le Mai Hoang K, Thi Tran H et al (2018) Identification of a biosynthetic gene cluster for the polyene macrolactam sceliphrolactam in a Streptomyces strain isolated from mangrove sediment. Sci. Rep 8:1594

    PubMed  PubMed Central  Google Scholar 

  • Madala NE, Piater LA, Steenkamp PA, Dubery IA (2014) Multivariate statistical models of metabolomic data reveals different metabolite distribution patterns in isonitrosoacetophenone-elicited Nicotiana tabacum and Sorghum bicolor cells. SpringerPlus 3:254

    PubMed  PubMed Central  Google Scholar 

  • Mo J, Wang S, Zhang W, Li C, Deng Z, Zhang L, Qu X (2019) Efficient editing DNA regions with high sequence identity in actinomycetal genomes by a CRISPR-Cas9 system. Synth Syst Biotechnol 4:86–91

    PubMed  PubMed Central  Google Scholar 

  • Mohana NC, Rao HY, Rakshith D, Mithun PR, Nuthan BR, Satish S (2018) Omics based approach for biodiscovery of microbial natural products in antibiotic resistance era. Genet Eng Biotechnol 16:1–8

    Google Scholar 

  • Mohimani H, Liu WT, Mylne JS, Poth AG, Colgrave ML, Tran D, Selsted ME, Dorrestein PC, Pevzner PA (2011) Cycloquest: identification of cyclopeptides via database search of their mass spectra against genome databases. J Proteome Res 10:4505–4512

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mohimani H, Liu WT, Kersten RD, Moore BS, Dorrestein PC, Pevzner PA (2014) NRPquest: coupling mass spectrometry and genome mining for nonribosomal peptide discovery. J Nat Prod 77:1902–1909

    CAS  PubMed  PubMed Central  Google Scholar 

  • Nguyen QT, Merlo ME, Medema MH, Jankevics A, Breitling R, Takano E (2012) Metabolomics methods for the synthetic biology of secondary metabolism. FEBS Lett 586:2177–2183

    CAS  PubMed  Google Scholar 

  • Nielsen ML, Isbrandt T, Rasmussen KB, Thrane U, Hoof JB, Larsen TO, Mortensen UH (2017) Genes linked to production of secondary metabolites in Talaromyces atroroseus revealed using CRISPR-Cas9. PLoS ONE 12:e0169712

    PubMed  PubMed Central  Google Scholar 

  • Palazzotto E, Weber T (2018) Omics and multi-omics approaches to study the biosynthesis of secondary metabolites in microorganisms. Curr Opin Biotechnol 45:109–116

    CAS  Google Scholar 

  • Palazzotto E, Tong Y, Lee SY, Weber T (2019) Synthetic biology and metabolic engineering of actinomycetes for natural product discovery. Biotechnol Adv 37:107366

    PubMed  Google Scholar 

  • Pohl C, Kiel JAKW, Driessen AJM, Bovenberg RAL, Nygard Y (2016) CRISPR/Cas9 based genome editing of Penicillium chrysogenum. ACS Synth Biol 5:754–764

    CAS  PubMed  Google Scholar 

  • Qin Z, Munnoch JT, Devine R, Holmes NA, Seipke RF, Wilkinson KA, Wilkinson B, Hutchings MI (2017) Formicamycins, antibacterial polyketides produced by Streptomyces formicae isolated from African Tetraponera plant-ants. Chem Sci 8:3218–3227

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ramell AM, Rodriguez MP, Alay A, Tulipani S, Urpi-sarda M, Sanchez-Pla A, Andres-Lacueva C (2018) Evaluation and comparison of bioinformatic tools for the enrichment analysis of metabolomics data. BMC Bioinform 19:1

    Google Scholar 

  • Rutledge PJ, Challis GL (2015) Discovery of microbial natural products by activation of silent biosynthetic gene clusters. Nat Rev Microbiol 13:509–523

    CAS  PubMed  Google Scholar 

  • Saini DK, Chakdar H, Pabbi S, Shukla P (2019) Enhancing production of microalgal biopigments through metabolic and genetic engineering. Crit Rev Food Sci Nutr 1:1–15

    Google Scholar 

  • Sander JD, Joung JK (2014) CRISPR-Cas systems for editing, regulating and targeting genomes. Nat Biotechnol 32:347–355

    CAS  PubMed  PubMed Central  Google Scholar 

  • Schrimpe-Rutledge AC, Sherrod SD, McLean JA (2018) Improving the discovery of secondary metabolite natural products using ion mobility–mass spectrometry. Curr Opin Chem Biol 42:160–166

    CAS  PubMed  Google Scholar 

  • Seyedsayamdost MR (2019) Toward a global picture of bacterial secondary metabolism. J Ind Microbiol Biotechnol 46:301–311

    CAS  PubMed  PubMed Central  Google Scholar 

  • Shao Z, Rao G, Li C, Abil Z, Luo Y, Zhao H (2013) Refactoring the silent spectinabilin gene cluster using a plug-and-play scaffold. ACS Synth Biol 2:662–669

    CAS  PubMed  PubMed Central  Google Scholar 

  • Singh BP, Rateb M, Rodriguez-Couto S, Polizeli MDLTD, Li WJ (2019) Microbial secondary metabolites: recent developments and technological challenges. Front Microbiol 10000:10000. https://doi.org/10.3389/fmicb.2019.00914

    Article  Google Scholar 

  • Tao W, Yang A, Deng Z, Sun Y (2018) CRISPR/Cas9-based editing of Streptomyces for discovery, characterization and production of natural products. Front Microbiol 9:1660

    PubMed  PubMed Central  Google Scholar 

  • Tawfik N, Tawfike ŮA, Abdo R, Abbott G, Abdelmohsen UR, Edrada-Ebel R, Haggag E (2017) Metabolomics and bioactivity guided isolation of secondary metabolites from the endophytic fungus Chaetomium sp. J Adv Pharm Res 1:66–74

    Google Scholar 

  • Tawfike AF, Romli M, Clements C, Abbott G, Young L, Schumacher M, Diederich M, Farag M, Edrada-Ebel R (2019) Isolation of anticancer and anti-trypanosome secondary metabolites from the endophytic fungus Aspergillus flocculus via bioactivity guided isolation and MS based metabolomics. J Chromatogr B 1106:71–83

    Google Scholar 

  • Tong Y, Charusanti P, Zhang L, Weber T, Lee SY (2015) CRISPR-Cas9 based engineering of actinomycetal genomes. ACS Synth Biol 4:1020–1029

    CAS  PubMed  Google Scholar 

  • Tong Y, Weber T, Lee SY (2019) CRISPR/Cas-based genome engineering in natural product discovery. Nat Prod Rep 36:1262–1280

    CAS  PubMed  Google Scholar 

  • Tran PN, Yen MR, Chiang CY, Lin HC, Chen PY (2019) Detecting and prioritizing biosynthetic gene clusters for bioactive compounds in bacteria and fungi. Applied Microbiol Biotechnol 103:3277–3287

    CAS  Google Scholar 

  • Valayil JM (2016) Activation of microbial silent gene clusters: genomics driven drug discovery approaches. Biochem Anal Biochem 5:276

    Google Scholar 

  • Wen B, Mei Z, Zeng C, Liu S (2017) metaX: a flexible and comprehensive software for processing metabolomics data. BMC Bioinformatics 18:183

    PubMed  PubMed Central  Google Scholar 

  • Wietz M, Mansson M, Gotfredsen CH, Larsen TO, Gram L (2010) Antibacterial compounds from marine Vibrionaceae isolated on a global expedition. Mar Drugs 8:2946–2960

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wishart DS (2016) Emerging applications of metabolomics in drug discovery and precision medicine. Nat Rev Drug Discov 15:473

    CAS  PubMed  Google Scholar 

  • Wu C, Kim HK, van Wezel GP, Choi YH (2015) Metabolomics in the natural products field—a gateway to novel antibiotics. Drug Discov Today Technol 13:11–17

    PubMed  Google Scholar 

  • Wu C, Zhu H, van Wezel GP, Choi YH (2016) Metabolomics-guided analysis of isocoumarin production by Streptomyces species MBT76 and biotransformation of flavonoids and phenylpropanoids. Metabolomics 12:90

    PubMed  PubMed Central  Google Scholar 

  • Wu C, Du C, Gubbens J, Choi YH, van Wezel GP (2018) Metabolomics-driven discovery of a prenylated isatin antibiotic produced by Streptomyces species MBT28. J Nat Prod 78:2355–2363

    Google Scholar 

  • Xia J, Psychogios N, Young N, Wishart DS (2009) MetaboAnalyst: a web server for metabolomic data analysis and interpretation. Nucleic Acids Res 37(suppl_2):W652–W660.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Xu M, Wright GD (2019) Heterologous expression-facilitated natural products’ discovery in actinomycetes. J Ind Microbiol Biotechnol 46:415–431

    CAS  PubMed  Google Scholar 

  • Yamanaka K, Reynolds KA, Kersten RD, Ryan KS, Gonzalez DJ, Nizet V et al (2014) Direct cloning and refactoring of a silent lipopeptide biosynthetic gene cluster yields the antibiotic taromycin A. Proc Natl Acad Sci USA 111:1957–1962

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zebec Z, Scrutton NS (2018) Genome editing for the production of natural products in Escherichia coli. Adv Biosyst 2:1800056

    Google Scholar 

  • Zhang MM, Wang Y, Ang EL, Zhao H (2016) Engineering microbial hosts for production of bacterial natural products. Nat Prod Rep 33:963–987

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang MM, Wong FT, Wang Y, Luo S, Lim YH, Heng E, Yeo WL, Cobb RE, Enghiad B, Ang EL, Zhao H (2017) CRISPR–Cas9 strategy for activation of silent Streptomyces biosynthetic gene clusters. Nat Chem Biol 13:607

    CAS  Google Scholar 

  • Zhang MM, Qiao Y, Ang EL, Zhao H (2018) Using natural products for drug discovery: the impact of the genomics era. Expert Opin Drug Discov 12:475–487

    Google Scholar 

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Acknowledgements

The author, Babita Sharma acknowledges Maharshi Dayanand University, Rohtak, India for providing University Research Fellowship (URS). PS acknowledges the infrastructural support from Department of Science and Technology, New Delhi, Govt. of India, through FIST grant (Grant No. 1196 SR/FST/LS-I/ 2017/4) and Department of Biotechnology, Government of India (Grant No. BT/PR27437/BCE/8/1433/2018) is duly acknowledged.

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Sinha, R., Sharma, B., Dangi, A.K. et al. Recent metabolomics and gene editing approaches for synthesis of microbial secondary metabolites for drug discovery and development. World J Microbiol Biotechnol 35, 166 (2019). https://doi.org/10.1007/s11274-019-2746-2

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