Laboratory-scale production of 13C-labeled lycopene and phytoene by bioengineered Escherichia coli.
Laboratory-scale production of 13C-labeled lycopene and phytoene by bioengineered Escherichia coli.
复制标题
通过生物工程大肠杆菌在实验室规模生产 13C 标记的番茄红素和八氢番茄红素。
DOI：
10.1021/jf202599z
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发表时间：
2011
影响因子：
6.1
通讯作者：
ErdmanJr,JohnW
中科院分区：
文献类型：
--
作者：
Lu,Chi-Hua;Choi,Jin-Ho;EngelmannMoran,Nancy;Jin,Yong-Su;ErdmanJr,JohnW
食用番茄制品与降低心血管疾病和癌症等慢性疾病的风险有关，因此番茄红素、八氢番茄红素和六氢番茄红素等番茄类胡萝卜素的生物学功能正在被研究。为了研究这些类胡萝卜素的吸收、分布、代谢和排泄，对生物工程大肠杆菌模型进行了评估，以实验室规模生产稳定同位素标记的类胡萝卜素。将来自成团肠杆菌的类胡萝卜素生物合成基因引入BL21Star(DE3)菌株中以产生番茄红素。超过96%的累积番茄红素处于全转化状态，并且通过13 C-葡萄糖给药，分子高度富集13 C。此外，使用易错 PCR 来破坏八氢番茄红素去饱和酶 (crtI) 功能并创建八氢番茄红素积累菌株，该菌株还被发现可以维持八氢番茄红素合酶 (crtB) 的转录。当13 C-葡萄糖是唯一碳源时，八氢番茄红素分子也高度富集13 C。该生产模型的开发将为类胡萝卜素研究人员提供标记示踪材料的来源，以进一步研究这些类胡萝卜素的代谢和生物功能。
Consumption of tomato products has been associated with decreased risks of chronic diseases such as cardiovascular disease and cancer, and therefore the biological functions of tomato carotenoids such as lycopene, phytoene, and phytofluene are being investigated. To study the absorption, distribution, metabolism, and excretion of these carotenoids, a bioengineered Escherichia coli model was evaluated for laboratory-scale production of stable isotope-labeled carotenoids. Carotenoid biosynthetic genes from Enterobacter agglomerans were introduced into the BL21Star(DE3) strain to yield lycopene. Over 96% of accumulated lycopene was in theall-transform, and the molecules were highly enriched with13C by13C-glucose dosing. In addition, error-prone PCR was used to disrupt phytoene desaturase (crtI) function and create a phytoene-accumulating strain, which was also found to maintain the transcription of phytoene synthase (crtB). Phytoene molecules were also highly enriched with13C when the13C-glucose was the only carbon source. The development of this production model will provide carotenoid researchers a source of labeled tracer materials to further investigate the metabolism and biological functions of these carotenoids.