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Benefits
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Bioflavonoids are a class of water-soluble plant pigments believed to have favorable medicinal qualities (hence their extensive use in nutraceuticals). However, the standard method for obtaining bioflavonoids—extraction from fruits such as blueberries and grapes—has disadvantages. These limitations are linked to the availability of the source fruits and the ability to consistently and selectively extract the desired compounds.
This patent-pending in vitro synthesis technology successfully addresses those disadvantages. It enables bioflavonoids to be available year-round, easily isolated, and consistently produced with higher yields than from natural fruits. The in vitro cell culture system can be manipulated to alter the composition of the bioflavonoids produced, providing a means to obtain large quantities of rare compounds. In addition, the bioflavonoids can be radiolabeled for use in bioavailability studies.
More information about this technology is presented below. For more information about this licensing opportunity, please contact us by phone or email: (919) 249-0327,
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The University’s in vitro bioflavonoid technology, which uses ohelo cell culture production, has several advantages over conventional fruit-extraction methods:
- Year-round availability: Because they are produced in vitro, these bioflavonoids are available year-round.
- Consistent quality: Studies demonstrate that the in vitro–produced bioflavonoids are of a consistent quality.
- Selectable: Users can select for production of unique compounds, such as epigalloCG, chlorogeic acid, catechin, ferulic acid, quercetrin, and epicatechin.
- Ability to label: Bioflavonoids produced using this technology can be labeled, allowing recovery and purification.
- High yield: Tests show that this in vitro production technology can yield significantly higher quantities of bioflavonoids than via most standard fruit extraction methods (see Figure 1).
FIGURE 1: Total Bioflavonoid Quantities Produced by Fruit Extraction and In Vitro Culture
The University’s in vitro production method with Ohelo cells generates significantly higher amounts of various bioflavonoids than other fruit extraction methods.
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Nutritional Supplements
This technology provides year-round availability of bioflavonoids that are primarily extracted from conventionally grown fruits, which are subject to variable climate conditions. This allows for an uninterrupted supply of bioflavonoids for the production of vitamins, herbal/botanical extracts, sports bars and beverages, and other products.
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Bioavailability Markers
The bioflavonoids produced by cell culture can be radioactively labeled and used as markers of bioavailability to track their metabolism.
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Compound Identification
This technology can produce compounds that can be used as a source of standards to be used for comparison to identify unknown compounds (e.g., to identify "active" ingredients).
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Food Coloring
Compounds made with this cell culture technology are more consistently produced than those extracted from fruits, yielding a reliable source for natural food dyes and gels. |
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How it Works
Vaccinium pahalae is a native of Hawaii that is a relative of blueberries and cranberries. Much anecdotal evidence suggests that such berry fruits have health benefits. But because extracting components and conducting bioactivity assays are difficult, little scientific evidence of these health benefits exists. The University’s technology will make bioactivity assays significantly easier to perform.
Cell cultures of V. pahalae were initiated from stable shoot microcultures. Unpigmented cell cultures were kept in the dark on a rotary shaker. These cell cultures were maintained by subculturing into fresh medium at 7-day intervals. The suspension medium was composed of Woody Plant Medium major and minor salts, rose vitamins, sucrose, polyvinylpyrrolidone, and specific plant growth regulators. In order to establish pigmented cell cultures, cells from the cultures incubated in the dark were transferred into a color-inducing medium. This medium contained a higher concentration of sucrose and the plant growth regulator benzyladenine instead of kinetin. These cell cultures were incubated under 100 micromol m-2 s-1 irradiance provided by cool white fluorescent lights. Cell cultures growing in the light were subcultured every 12 to 14 days.
Cell cultures were filtered then extracted with 70% acetone. After removing the acetone, the aqueous solution was freeze-dried to yield a red solid. Extracts may be prepared in a similar manner from fruits. Samples of the extract were mixed with silica gel 60, air dried, then loaded on a column with silica gel 60. The column was washed with petroleum ether before and after loading the extract. Fractions were eluted with ethyl acetate; increasing amounts of methanol-water were added up to 100% methanol-water. These fractions were concentrated and lyophilized, and selected fractions were further fractionated by chromatography on either silica gel or Sephadex LH-20.
Inventors currently have scaled up the technology to a 12-liter bioreactor with a 10-liter working volume.
Patent Applications
- Methods for Isolation of Proanthocyanidins from Flavonoid-Producing Cell Culture
Published Papers
- "Isolation of oligomeric proanthocyanidins from flavonoid-producing cell cultures," by F.E. Kandil, L. Song, J.M. Pezzuto, K. Marley, D.S. Seigler, and M.A.L. Smith. Published in In Vitro Cellular and Developmental Biology – Plant, 2000, 36(6):492–500.
- "Anthocyanin production from Vaccinium pahalae: limitations of the physical microenvironment," by J.E. Meyer, M.-F. Pépin, M.A.L. Smith. Published in Journal of Biotechnology, 2002, 93(1):45-57.
- "An in vitro approach to investigate medicinal chemical synthesis by three herbal plants," by M.A.L. Smith, H. Kobayashi, M. Gawienowski, and D.P. Briskin. Published in Plant Cell, Tissue and Organ Culture (in press).
- "Composition of a chemopreventive proanthocyanidin-rich fraction from cranberry fruits responsible for the inhibition of 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced ornithine decarboxylase (ODC) activity" by F.E. Kandil, M.A.L. Smith, R.B. Rogers, M-F Pépin, L.L. Song, J.M. Pezzuto, and D.S. Seigler. Published in the Journal of Agriculture and Food Chemistry, 2002, 50(5):1063–1069.
Technical Presentations
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The University of Illinois at Urbana–Champaign is offering its in vitro bioflavonoid production technology for license by and/or joint development with commercial companies, universities, or government labs. Provisional U.S. patents have been applied for, and this technology is fully owned by the University.
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