Abstract
Triterpene saponins of the genus Quillaja (Quillajaceae) are known for their immunoadjuvant, hypocholesterolemic, and anti-inflammatory activity. Plant cell cultures are useful for the study of saponin metabolism and industrial production of these bioactive compounds. While structurally related phytosterols are primary metabolites essential to growth and development, saponins are responsive to pathogen and abiotic stress, fulfilling roles in plant specialized metabolism. For cell culture production of saponins, phytosterols may be considered a competing pathway which relies on a common pool of cytosolic isoprenoid precursors.
Understanding the metabolic allocation of resources between these two related pathways is key to maximizing saponin production in in vitro production systems. Sterols and saponins naturally occur in multiple conjugated forms, which complicate separation and quantification. The acid hydrolysis of conjugated sterols and saponins to their free forms is a useful technique to simplify their analysis by gas chromatography. Here we provide the workflow for the quantification of free sterols and sapogenins in cell cultures of Quillaja brasiliensis .
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References
Lara JA, Burciaga-Monge A, Chávez A et al (2018) Identification and characterization of sterol acyltransferases responsible for steryl ester biosynthesis in tomato. Front Plant Sci 9:588
Reichert CL, Salminen H, Weiss J (2019) Quillaja saponin characteristics and functional properties. Annu Rev Food Sci Tech 10:43–73
Osbourn A, Reed J (2019) Metabolic engineering. Patent cooperation treaty EP2018/086430, application WO 2019/122259 A1, 20 Dec 2018
Magedans YV, Yendo AC, de Costa F et al (2019) Foamy matters: an update on Quillaja saponins and their use as immunoadjuvants. Future Med Chem 11:1485–1499
Kensil CR, Patel U, Lennick M et al (1991) Separation and characterization of saponins with adjuvant activity from Quillaja saponaria Molina cortex. J Immunol 146:431–437
Yendo ACA, de Costa F, Cibulski SP et al (2016) A rabies vaccine adjuvanted with saponins from leaves of the soap tree (Quillaja brasiliensis) induces specific immune responses and protects against lethal challenge. Vaccine 34:2305–2311
Santos FN, Borja-Cabrera GP, Miyashiro L et al (2007) Immunotherapy against experimental canine visceral leishmaniasis with the saponin enriched-Leishmune® vaccine. Vaccine 25:6176–6190
James SF, Chahine EB, Sucher AJ, Hanna C (2018) Shingrix: the new adjuvanted recombinant herpes zoster vaccine. Ann Pharmacother 52:673–680
Guerra Mendoza Y, Garric E, Leach A et al (2019) Safety profile of the RTS, S/AS01 malaria vaccine in infants and children: additional data from a phase III randomized controlled trial in sub-Saharan Africa. Hum Vaccin Immunother 15:2386–2398
Arslan I (2020) Quillaic acid–containing saponin-based immunoadjuvants trigger early immune responses. Rev Bras Farmacogn 30:467–473
Roberts A, Lamirande EW, Vogel L et al (2010) Immunogenicity and protective efficacy in mice and hamsters of a β-propiolactone inactivated whole virus SARS-CoV vaccine. Viral Immunol 23:509–519
Gupta T, Gupta SK (2020) Potential adjuvants for the development of a SARS-CoV-2 vaccine based on experimental results from similar coronaviruses. Int Immunopharmacol 86:106717
Sharma R, Palanisamy A, Dhama K et al (2020) Exploring the possible use of saponin adjuvants in COVID-19 vaccine. Hum Vaccin Immunother 16:2944–2953
da Silva MYV, Phillips MA, Fett-Neto AG (2020) Production of plant bioactive triterpenoid saponins: from metabolites to genes and back. Phytochem Rev 20:461–482
Valitova JN, Sulkarnayeva AG, Minibayeva FV (2016) Plant sterols: diversity, biosynthesis, and physiological functions. Biochem Mosc 81:819–834
Ferrer A, Altabella T, Arró M, Boronat A (2017) Emerging roles for conjugated sterols in plants. Prog Lipid Res 67:27–37
Rogowska A, Szakiel A (2020) The role of sterols in plant response to abiotic stress. Phytochem Rev 19:1525–1538
Planas-Riverola A, Gupta A, Betegón-Putze I et al (2019) Brassinosteroid signaling in plant development and adaptation to stress. Development 146:dev151894
Osbourn A, Goss RJM, Field RA (2011) The saponins—polar isoprenoids with important and diverse biological activities. Nat Prod Rep 28:1261–1268
Yang C-R, Zhang Y, Jacob MR et al (2006) Antifungal activity of C-27 steroidal saponins. Antimicrob Agents Chemother 50:1710–1714
Meesapyodsuk D, Balsevich J, Reed DW et al (2007) Saponin biosynthesis in Saponaria vaccaria. cDNAs encoding β-amyrin synthase and a triterpene carboxylic acid glucosyltransferase. Plant Physiol 143:959–969
Assimopoulou A, Papageorgiou V (2005) GC-MS analysis of penta-and tetra-cyclic triterpenes from resins of Pistacia species. Part I. Pistacia lentiscus var. Chia. Biomed Chromatogr 19:285–311
Lai C, Li S, Yu H et al (2006) A rapid HPLC–ESI-MS/MS for qualitative and quantitative analysis of saponins in “XUESETONG” injection. J Pharm Biomed Anal 40:669–678
Christie W, Han X (2010) Lipid analysis. The Oily Press, Bridgewater
Abidi S (2001) Chromatographic analysis of plant sterols in foods and vegetable oils. J Chromatogr A 935:173–201
Moreau RA, Nyström L, Whitaker BD et al (2018) Phytosterols and their derivatives: structural diversity, distribution, metabolism, analysis, and health-promoting uses. Prog Lipid Res 70:35–61
Flores-Sánchez IJ, Ortega-López J, Montes-Horcasitas M d C et al (2002) Biosynthesis of sterols and triterpenes in cell suspension cultures of Uncaria tomentosa. Plant Cell Physiol 43:1502–1509
Moreau RA, Whitaker BD, Hicks KB (2002) Phytosterols, phytostanols, and their conjugates in foods: structural diversity, quantitative analysis, and health-promoting uses. Prog Lipid Res 41:457–500
Mustafa NR, De Winter W, Van Iren F et al (2011) Initiation, growth and cryopreservation of plant cell suspension cultures. Nat Protoc 6:715–742
Yendo AC, de Costa F, Kauffmann C et al (2017) Purification of an immunoadjuvant saponin fraction from Quillaja brasiliensis leaves by reversed-phase silica gel chromatography. In: Fox CB (ed) Vaccine Adjuvants, Methods in molecular biology, vol 1494. Springer, New York, pp 87–93
Tava A, Biazzi E, Mella M et al (2017) Artefact formation during acid hydrolysis of saponins from Medicago spp. Phytochemistry 138:116–127
Fleck JD, de Costa F, Yendo AC et al (2013) Determination of new immunoadjuvant saponin named QB-90, and analysis of its organ-specific distribution in Quillaja brasiliensis by HPLC. Nat Prod Res 27:907–910
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Magedans, Y.V.S., Phillips, M.A. (2022). Soapbark Triterpenes: Quillaja brasiliensis Cell Culture Sapogenin and Free Sterol Analysis by GCMS . In: Fett-Neto, A.G. (eds) Plant Secondary Metabolism Engineering. Methods in Molecular Biology, vol 2469. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2185-1_10
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DOI: https://doi.org/10.1007/978-1-0716-2185-1_10
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