Dataset Information

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.

SUBMITTER: Smith JW

PROVIDER: S-EPMC5384503 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

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Carrot solution culture bioproduction of uniformly labeled <sup>13</sup>C-lutein and in vivo dosing in non-human primates.

Smith Joshua W JW Rogers Randy B RB Jeon Sookyoung S Rubakhin Stanislav S SS Wang Lin L Sweedler Jonathan V JV Neuringer Martha M Kuchan Matthew J MJ Erdman John W JW

Experimental biology and medicine (Maywood, N.J.) 20161023 3

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 ...[more]

PMID: 27798119

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Project description:BackgroundStable isotopically labelled organisms have found wide application in life science research including plant physiology, plant stress and defense as well as metabolism related sciences. Therefore, the reproducible production of plant material enriched with stable isotopes such as 13C and 15N is of considerable interest. A high degree of enrichment (> 96 atom %) with a uniformly distributed isotope (global labelling) is accomplished by a continuous substrate supply during plant growth/cultivation. In the case of plants, 13C-labelling can be achieved by growth in 13CO2(g) atmosphere while global 15N-labelling needs 15N- containing salts in the watering/nutrient solution. Here, we present a method for the preparation of 13C and 15N-labelled plants by the use of closed growth chambers and hydroponic nutrient supply. The method is exemplified with durum wheat.ResultsIn total, 330 g of globally 13C- and 295 g of 15N-labelled Triticum durum wheat was produced during 87 cultivation days. For this, a total of 3.88 mol of 13CO2(g) and 58 mmol of 15N were consumed. The degree of enrichment was determined by LC-HRMS and ranged between 96 and 98 atom % for 13C and 95-99 atom % for 15N, respectively. Additionally, the isotopically labelled plant extracts were successfully used for metabolome-wide internal standardisation of native T.durum plants. Application of an isotope-assisted LC-HRMS workflow enabled the detection of 652 truly wheat-derived metabolites out of which 143 contain N.ConclusionA reproducible cultivation which makes use of climate chambers and hydroponics was successfully adapted to produce highly enriched, uniformly 13C- and 15N-labelled wheat. The obtained plant material is suitable to be used in all kinds of isotope-assisted research. The described technical equipment and protocol can easily be applied to other plants to produce 13C-enriched biological samples when the necessary specific adaptations e.g. temperature and light regime, as well as nutrient supply are considered. Additionally, the 15N-labelling method can also be carried out under regular glasshouse conditions without the need for customised atmosphere.

Dataset Information

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

Publications

Carrot solution culture bioproduction of uniformly labeled <sup>13</sup>C-lutein and in vivo dosing in non-human primates.

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

Publications

Carrot solution culture bioproduction of uniformly labeled <sup>13</sup>C-lutein and in vivo dosing in non-human primates.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

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