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Production of Uniformly Labeled 13 C‐Lutein and 13 C‐α‐Tocopherol in vitro Using Carrot Cell Suspension Culture
Author(s) -
Smith Joshua,
Rogers Randy,
Jeon Sookyoung,
Rubakhin Stanislav,
Wang Lin,
Sweedler Jonathan,
Kuchan Matthew,
Erdman John
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.604.3
Subject(s) - lutein , daucus carota , chemistry , tocopherol , lookup table , high performance liquid chromatography , chromatography , biochemistry , antioxidant , vitamin e , carotenoid , botany , biology , computer science , programming language
The xanthophyll lutein (Lut) is present in high concentrations in primate retinae, protecting against photooxidation and enhancing visual performance. Interestingly, in the elderly, diets high in Lut slow cognitive decline and circulating levels of both Lut and the lipid‐soluble antioxidant α‐tocopherol (αT) are inversely associated with cognitive impairment. However, Lut deposition, metabolism, and function in the brain are poorly understood, and cost‐effective isotopically labeled Lut tracers are not commercially available. The goal of this work, therefore, is to establish a system for efficient bioproduction of uniformly labeled 13 C‐Lut and 13 C‐αT. Seven carrot ( Daucus carota ) cultivars were germinated, separated into tissue subtypes, dedifferentiated into solid callus, and initiated in liquid suspension culture. Conditions for production of Lut and αT in suspension culture were optimized and uniformly labeled 13 C‐glucose was provided as the sole media carbon source for four serial growth cycles. Lut and αT yields were 2.6 and 3.3 μg/g cells, respectively, as measured by reverse‐phase HPLC. Cell extracts were fractionated by preparatory reverse‐phase HPLC and analyzed for 13 C enrichment by mass spectrometry. 58% of Lut and 50% of αT in carrot extract fractions were uniformly labeled with 13 C. These tracers can be utilized in studies of Lut and αT in the brain, enhancing our understanding of their function in cognition.