Engelmann Nancy J, Rogers Randy B, Lila Mary Ann, Erdman John W
Division of Nutritional Sciences, University of Illinois, Urbana/Champaign, Urbana, Illinois 61801, USA.
J Agric Food Chem. 2009 Jun 10;57(11):4614-9. doi: 10.1021/jf803905d.
Progress in learning about underlying carotenoid bioactivity mechanisms has been limited because of the lack of commercially available radiolabeled lycopene (LYC), phytoene (PE), and phytofluene (PF). Tomato ( Solanum lycopersicum cv. VFNT cherry) cell cultures have been treated to produce [(14)C]-PE and PF but with relatively low yields. To increase carotenoid production, two bleaching herbicides were administered during the culture incubation, 2-(4-chlorophenyl-thio)triethylamine and norflurazon, separately or in combination to produce varying ratios of PE, PF, and LYC. Treatment with both herbicides resulted in optimal production of all three carotenoids. Subsequently, cultures were incubated in [(14)C]-sucrose-containing media to produce labeled LYC, PE, and PF. Adding [(14)C]-sucrose on day 1 of the 14 day culture incubation cycle to norflurazon-treated cultures led to a small increase in labeling efficiency compared to adding it on day 7. Improved culture conditions efficiently provided sufficient (14)C-carotenoids for future cell culture and animal metabolic tracking studies.
由于缺乏市售的放射性标记番茄红素(LYC)、八氢番茄红素(PE)和六氢番茄红素(PF),关于潜在类胡萝卜素生物活性机制的研究进展有限。番茄(Solanum lycopersicum cv. VFNT cherry)细胞培养物已被处理以产生[(14)C]-PE和PF,但产量相对较低。为了提高类胡萝卜素的产量,在培养孵育期间分别或联合施用两种漂白除草剂,即2-(4-氯苯基硫代)三乙胺和氟草敏,以产生不同比例的PE、PF和LYC。两种除草剂处理均导致所有三种类胡萝卜素的最佳产量。随后,将培养物在含[(14)C]-蔗糖的培养基中孵育,以产生标记的LYC、PE和PF。与在第7天添加[(14)C]-蔗糖相比,在14天培养孵育周期的第1天向氟草敏处理的培养物中添加[(14)C]-蔗糖导致标记效率略有提高。改进的培养条件有效地提供了足够的(14)C-类胡萝卜素,用于未来的细胞培养和动物代谢追踪研究。