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Production of [ 13 C]‐lycopene from high lycopene tomato cell suspension cultures
Author(s) -
Engelmann Nancy J.,
Rogers Randy B.,
Rupassara S. Indu,
Garlick Peter J.,
Lila Mary Ann,
Erdman John W.
Publication year - 2010
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.24.1_supplement.539.6
Subject(s) - chemistry , lycopene , phytoene , carotenoid , food science , enzyme , suspension culture , biochemistry , nuclear chemistry , cell culture , biology , genetics
The goal of this work was to use tomato cell suspension cultures treated with enzyme inhibitors and grown with [ 13 C 6 ]‐glucose as a carbon source to produce labeled carotenoids (CAR) for human metabolism research. To promote CAR accumulation, the high lycopene (LYC) tomato cell suspension line ‘Ailsa Craig’ hp‐1 was grown with carotenogenic enzyme inhibitors norflurazon (NORF), 2‐(4‐chlorophenyl‐thio) triethylamine (CPTA), both herbicides (BOTH), or neither (CONT). Significantly greater combined CAR [phytoene (PE), phytofluene (PF), and LYC] yields were obtained with the NORF, CPTA, and BOTH treatments (5.2 ± 0.7, 3.6 ± 0.4, 4.2 ± 0.4 mg/L, respectively) than CONT (0.3 ± 0.1 mg/L). Different PE, PF, and LYC profiles were obtained where CPTA led to 0.26 ± 0.02, 0.14 ± 0.02, and 3.18 ± 0.45 mg/L; NORF led to 4.96 ± 0.68, 0.20 ± 0.06, and 0.06 ± 0.02 mg/L; BOTH led to 2.98 ± 0.14, 0.18 ± 0.05, and 1.05 ± 0.36 mg/L; CONT led to 0.18 ± 0.09, 0.02 ± 0.00, and 0.06 ± 0.01 mg/L, respectively. To biolabel LYC the hp‐1 cell line was grown in medium containing 30 g/L [ 13 C 6 ]‐glucose and CPTA. LYC, PE, and PF yields from labeled cultures were 2.5 ± 0.1, 0.3 ± 0.0, 0.1 ± 0.0 mg/L, respectively. Mass isotopomer distribution analysis for LYC [ 13 C]‐enrichment from a pilot labeling run was performed using LC‐MS/MS, and 82% of the LYC molecules had at least 34 of 40 carbons labeled, and the greatest percent (17%) was uniformly labeled. (NIH/NCI CA 112649‐01A1)