Carrot solution culture bioproduction of uniformly labeled 13C-lutein and in vivo dosing in non-human primates

Joshua W. Smith, Randy B. Rogers, Sookyoung Jeon, Stanislav S. Rubakhin, Lin Wang, Jonathan V. Sweedler, Martha Neuringer, Matthew J. Kuchan, John W. Erdman

    Research output: Contribution to journalArticle

    1 Citation (Scopus)

    Abstract

    Lutein is a xanthophyll abundant in nature and most commonly present in the human diet through consumption of leafy green vegetables. With zeaxanthin and meso-zeaxanthin, lutein is a component of the macular pigment of the retina, where it protects against photooxidation and age-related macular degeneration. Recent studies have suggested that lutein may positively impact cognition throughout the lifespan, but outside of the retina, the deposition, metabolism, and function(s) of lutein are poorly understood. Using a novel botanical cell culture system (Daucus carota), the present study aimed to produce a stable isotope lutein tracer for use in future investigations of dietary lutein distribution and metabolism. Carrot cultivars were initiated into liquid solution culture, lutein production conditions optimized, and uniformly labeled 13C-glucose was provided as the sole media carbon source for four serial growth cycles. Lutein yield was 2.58 ± 0.24 µg/g, and mass spectrometry confirmed high enrichment of 13C: 64.9% of lutein was uniformly labeled and 100% of lutein was labeled on at least 37 of 40 possible carbons. Purification of carrot extracts yielded a lutein dose of 1.92 mg with 96.0 ± 0.60% purity. 13C-lutein signals were detectable in hepatic extracts of an adult rhesus macaque monkey (Macaca mulatta) dosed with 13C-lutein, but not in hepatic samples collected from control animals. This novel botanical biofactory approach can be used to produce sufficient quantities of highly enriched and pure 13C-lutein doses for use in tracer studies investigating lutein distribution, metabolism, and function.

    Original languageEnglish (US)
    Pages (from-to)305-315
    Number of pages11
    JournalExperimental Biology and Medicine
    Volume242
    Issue number3
    DOIs
    StatePublished - Feb 1 2017

    Fingerprint

    Lutein
    Daucus carota
    Primates
    Macaca mulatta
    Metabolism
    Retina
    Carbon
    Xanthophylls
    Photooxidation
    Liver
    Vegetables
    Macular Degeneration
    Nutrition
    Cell culture

    Keywords

    • isotopic tracer
    • Lutein
    • mass spectrometry
    • non-human primate
    • nutrition
    • phytochemicals

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)

    Cite this

    Smith, J. W., Rogers, R. B., Jeon, S., Rubakhin, S. S., Wang, L., Sweedler, J. V., ... Erdman, J. W. (2017). Carrot solution culture bioproduction of uniformly labeled 13C-lutein and in vivo dosing in non-human primates. Experimental Biology and Medicine, 242(3), 305-315. https://doi.org/10.1177/1535370216675067

    Carrot solution culture bioproduction of uniformly labeled 13C-lutein and in vivo dosing in non-human primates. / Smith, Joshua W.; Rogers, Randy B.; Jeon, Sookyoung; Rubakhin, Stanislav S.; Wang, Lin; Sweedler, Jonathan V.; Neuringer, Martha; Kuchan, Matthew J.; Erdman, John W.

    In: Experimental Biology and Medicine, Vol. 242, No. 3, 01.02.2017, p. 305-315.

    Research output: Contribution to journalArticle

    Smith, JW, Rogers, RB, Jeon, S, Rubakhin, SS, Wang, L, Sweedler, JV, Neuringer, M, Kuchan, MJ & Erdman, JW 2017, 'Carrot solution culture bioproduction of uniformly labeled 13C-lutein and in vivo dosing in non-human primates', Experimental Biology and Medicine, vol. 242, no. 3, pp. 305-315. https://doi.org/10.1177/1535370216675067
    Smith, Joshua W. ; Rogers, Randy B. ; Jeon, Sookyoung ; Rubakhin, Stanislav S. ; Wang, Lin ; Sweedler, Jonathan V. ; Neuringer, Martha ; Kuchan, Matthew J. ; Erdman, John W. / Carrot solution culture bioproduction of uniformly labeled 13C-lutein and in vivo dosing in non-human primates. In: Experimental Biology and Medicine. 2017 ; Vol. 242, No. 3. pp. 305-315.
    @article{dd087943e10d440981aa4c300cbcf3a8,
    title = "Carrot solution culture bioproduction of uniformly labeled 13C-lutein and in vivo dosing in non-human primates",
    abstract = "Lutein is a xanthophyll abundant in nature and most commonly present in the human diet through consumption of leafy green vegetables. With zeaxanthin and meso-zeaxanthin, lutein is a component of the macular pigment of the retina, where it protects against photooxidation and age-related macular degeneration. Recent studies have suggested that lutein may positively impact cognition throughout the lifespan, but outside of the retina, the deposition, metabolism, and function(s) of lutein are poorly understood. Using a novel botanical cell culture system (Daucus carota), the present study aimed to produce a stable isotope lutein tracer for use in future investigations of dietary lutein distribution and metabolism. Carrot cultivars were initiated into liquid solution culture, lutein production conditions optimized, and uniformly labeled 13C-glucose was provided as the sole media carbon source for four serial growth cycles. Lutein yield was 2.58 ± 0.24 µg/g, and mass spectrometry confirmed high enrichment of 13C: 64.9{\%} of lutein was uniformly labeled and 100{\%} of lutein was labeled on at least 37 of 40 possible carbons. Purification of carrot extracts yielded a lutein dose of 1.92 mg with 96.0 ± 0.60{\%} purity. 13C-lutein signals were detectable in hepatic extracts of an adult rhesus macaque monkey (Macaca mulatta) dosed with 13C-lutein, but not in hepatic samples collected from control animals. This novel botanical biofactory approach can be used to produce sufficient quantities of highly enriched and pure 13C-lutein doses for use in tracer studies investigating lutein distribution, metabolism, and function.",
    keywords = "isotopic tracer, Lutein, mass spectrometry, non-human primate, nutrition, phytochemicals",
    author = "Smith, {Joshua W.} and Rogers, {Randy B.} and Sookyoung Jeon and Rubakhin, {Stanislav S.} and Lin Wang and Sweedler, {Jonathan V.} and Martha Neuringer and Kuchan, {Matthew J.} and Erdman, {John W.}",
    year = "2017",
    month = "2",
    day = "1",
    doi = "10.1177/1535370216675067",
    language = "English (US)",
    volume = "242",
    pages = "305--315",
    journal = "Experimental Biology and Medicine",
    issn = "1535-3702",
    publisher = "SAGE Publications Ltd",
    number = "3",

    }

    TY - JOUR

    T1 - Carrot solution culture bioproduction of uniformly labeled 13C-lutein and in vivo dosing in non-human primates

    AU - Smith, Joshua W.

    AU - Rogers, Randy B.

    AU - Jeon, Sookyoung

    AU - Rubakhin, Stanislav S.

    AU - Wang, Lin

    AU - Sweedler, Jonathan V.

    AU - Neuringer, Martha

    AU - Kuchan, Matthew J.

    AU - Erdman, John W.

    PY - 2017/2/1

    Y1 - 2017/2/1

    N2 - Lutein is a xanthophyll abundant in nature and most commonly present in the human diet through consumption of leafy green vegetables. With zeaxanthin and meso-zeaxanthin, lutein is a component of the macular pigment of the retina, where it protects against photooxidation and age-related macular degeneration. Recent studies have suggested that lutein may positively impact cognition throughout the lifespan, but outside of the retina, the deposition, metabolism, and function(s) of lutein are poorly understood. Using a novel botanical cell culture system (Daucus carota), the present study aimed to produce a stable isotope lutein tracer for use in future investigations of dietary lutein distribution and metabolism. Carrot cultivars were initiated into liquid solution culture, lutein production conditions optimized, and uniformly labeled 13C-glucose was provided as the sole media carbon source for four serial growth cycles. Lutein yield was 2.58 ± 0.24 µg/g, and mass spectrometry confirmed high enrichment of 13C: 64.9% of lutein was uniformly labeled and 100% of lutein was labeled on at least 37 of 40 possible carbons. Purification of carrot extracts yielded a lutein dose of 1.92 mg with 96.0 ± 0.60% purity. 13C-lutein signals were detectable in hepatic extracts of an adult rhesus macaque monkey (Macaca mulatta) dosed with 13C-lutein, but not in hepatic samples collected from control animals. This novel botanical biofactory approach can be used to produce sufficient quantities of highly enriched and pure 13C-lutein doses for use in tracer studies investigating lutein distribution, metabolism, and function.

    AB - Lutein is a xanthophyll abundant in nature and most commonly present in the human diet through consumption of leafy green vegetables. With zeaxanthin and meso-zeaxanthin, lutein is a component of the macular pigment of the retina, where it protects against photooxidation and age-related macular degeneration. Recent studies have suggested that lutein may positively impact cognition throughout the lifespan, but outside of the retina, the deposition, metabolism, and function(s) of lutein are poorly understood. Using a novel botanical cell culture system (Daucus carota), the present study aimed to produce a stable isotope lutein tracer for use in future investigations of dietary lutein distribution and metabolism. Carrot cultivars were initiated into liquid solution culture, lutein production conditions optimized, and uniformly labeled 13C-glucose was provided as the sole media carbon source for four serial growth cycles. Lutein yield was 2.58 ± 0.24 µg/g, and mass spectrometry confirmed high enrichment of 13C: 64.9% of lutein was uniformly labeled and 100% of lutein was labeled on at least 37 of 40 possible carbons. Purification of carrot extracts yielded a lutein dose of 1.92 mg with 96.0 ± 0.60% purity. 13C-lutein signals were detectable in hepatic extracts of an adult rhesus macaque monkey (Macaca mulatta) dosed with 13C-lutein, but not in hepatic samples collected from control animals. This novel botanical biofactory approach can be used to produce sufficient quantities of highly enriched and pure 13C-lutein doses for use in tracer studies investigating lutein distribution, metabolism, and function.

    KW - isotopic tracer

    KW - Lutein

    KW - mass spectrometry

    KW - non-human primate

    KW - nutrition

    KW - phytochemicals

    UR - http://www.scopus.com/inward/record.url?scp=85011636655&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=85011636655&partnerID=8YFLogxK

    U2 - 10.1177/1535370216675067

    DO - 10.1177/1535370216675067

    M3 - Article

    C2 - 27798119

    AN - SCOPUS:85011636655

    VL - 242

    SP - 305

    EP - 315

    JO - Experimental Biology and Medicine

    JF - Experimental Biology and Medicine

    SN - 1535-3702

    IS - 3

    ER -