Abstract
Antibiotic natural products from microbes are characterized by diverse and mostly complex chemical structures, which challenge their total chemical synthesis and make biotechnological production to the predominant production route. In order to reach these valuable compounds in the fermentation broth, sophisticated recovery methods are required, and a high degree of purity is essential for a thorough exploration of their beneficial properties in subsequent assays. The isolation and purification of natural products from microbial cultures is mainly based on the repeated application of extraction and chromatographic separation methods.
This chapter describes the general strategy of natural product recovery from microbial cultures, gives theoretical and practical insights to underlying methods—essentially compound extraction and preparative chromatography—and describes a specific methodical approach to isolate and purify the natural product fusarubin from the culture of the fungus Fusarium sp.
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References
Atanasov AG, Zotchev SB, Dirsch VM, International Natural Product Sciences T, Supuran CT (2021) Natural products in drug discovery: advances and opportunities. Nat Rev Drug Discov 20(3):200–216
Cragg GM, Newman DJ (2013) Natural products: a continuing source of novel drug leads. Biochim Biophys Acta 1830(6):3670–3695
Li JW, Vederas JC (2009) Drug discovery and natural products: end of an era or an endless frontier? Science 325(5937):161–165
Miethke M, Pieroni M, Weber T, Brönstrup M, Hammann P, Halby L, Arimondo PB, Glaser P, Aigle B, Bode HB, Moreira R, Li Y, Luzhetskyy A, Medema MH, Pernodet JL, Stadler M, Tormo JR, Genilloud O, Truman AW, Weissman KJ, Takano E, Sabatini S, Stegmann E, Brötz-Oesterhelt H, Wohlleben W, Seemann M, Empting M, Hirsch AKH, Loretz B, Lehr CM, Titz A, Herrmann J, Jaeger T, Alt S, Hesterkamp T, Winterhalter M, Schiefer A, Pfarr K, Hoerauf A, Graz H, Graz M, Lindvall M, Ramurthy S, Karlén A, van Dongen M, Petkovic H, Keller A, Peyrane F, Donadio S, Fraisse L, Piddock LJV, Gilbert IH, Moser HE, Müller R (2021) Towards the sustainable discovery and development of new antibiotics. Nat Rev Chem 5:726–749
Wilson BAP, Thornburg CC, Henrich CJ, Grkovic T, O'Keefe BR (2020) Creating and screening natural product libraries. Nat Prod Rep 37(7):893–918
Cannell RJP (1998) How to approach the isolation of a natural product. In: Cannell RJP (ed) Natural products isolation. Humana Press, Totowa, NJ, pp 1–51
Sarker SD, Nahar L (2012) An introduction to natural products isolation. In: Sarker SD, Nahar L (eds) Natural products isolation. Humana Press, Totowa, NJ, pp 1–25
Helaly SE, Kulik A, Zinecker H, Ramachandaran K, Tan GY, Imhoff JF, Süssmuth RD, Fiedler HP, Sabaratnam V (2012) Langkolide, a 32-membered macrolactone antibiotic produced by Streptomyces sp. Acta 3062. J Nat Prod 75(6):1018–1024
Nachtigall J, Kulik A, Helaly S, Bull AT, Goodfellow M, Asenjo JA, Maier A, Wiese J, Imhoff JF, Süssmuth RD, Fiedler HP (2011) Atacamycins A-C, 22-membered antitumor macrolactones produced by Streptomyces sp. C38. J Antibiot (Tokyo) 64(12):775–780
Fiedler H-P (1977) Isolierung von mikrobiellen Stoffwechselprodukten mit Hilfe von Adsorberharzen. Chemische Rundschau 30:2–3
Berlinck RGS, Crnkovic CM, Gubiani JR, Bernardi DI, Ióca LP, Quintana-Bulla JI (2021) The isolation of water-soluble natural products – challenges, strategies and perspectives. Nat Prod Rep
Arslanian RL, Parker CD, Wang PK, McIntire JR, Lau J, Starks C, Licari PJ (2002) Large-scale isolation and crystallization of epothilone D from Myxococcus xanthus cultures. J Nat Prod 65(4):570–572
Horosanskaia E, Triemer S, Seidel-Morgenstern A, Lorenz H (2019) Purification of artemisinin from the product solution of a semisynthetic reaction within a single crystallization step. Org Process Res Dev 23(9):2074–2079
Meyer V (2004) Präparative HPLC. In: Praxis der Hochleistungs-Flüssigchromatographie. Wiley GmbH & Co. KGaA, Weinheim, pp 285–296
Vailaya A, Horvath C (1998) Retention in reversed-phase chromatography: partition or adsorption? J Chromatogr A 829(1–2):1–27
Yoon KD, Chin YW, Kim J (2010) Centrifugal partition chromatography: application to natural products in 1994–2009. J Liq Chromatogr R T 33(9–12):1208–1254
Abdin AY, Yeboah P, Jacob C (2020) Chemical impurities: an epistemological riddle with serious side effects. Int J Environ Res Public Health 17(3)
CLSI (2019) Performance standards for antimicrobial susceptibility testing. CLSI guidline M100, 29th edn. Clinical and Laboratory Standards Institute, Wayne
Schafhauser T, Jahn L, Kirchner N, Kulik A, Flor L, Lang A, Caradec T, Fewer DP, Sivonen K, van Berkel WJH, Jacques P, Weber T, Gross H, van Pee KH, Wohlleben W, Ludwig-Müller J (2019) Antitumor astins originate from the fungal endophyte Cyanodermella asteris living within the medicinal plant Aster tataricus. Proc Natl Acad Sci U S A 116(52):26909–26917
Golay MJE (1958) Theory of chromatography in open and open tubular columns with round and rectangular cross sections. In: Desty DH (ed) Gas chromatography. Butterworths, London, pp 36–55
Kulik A, Fiedler H-P (1998) Some aspects of the purification of anthraquinone antibiotics by preparative reversed-phase liquid chromatography. J Chromatogr A 812:117–121
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Schafhauser, T., Kulik, A. (2023). Isolation and Purification of Natural Products from Microbial Cultures. In: Sass, P. (eds) Antibiotics. Methods in Molecular Biology, vol 2601. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2855-3_5
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DOI: https://doi.org/10.1007/978-1-0716-2855-3_5
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