Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Japan.
Research Institute for Production Development, Kyoto, Japan.
Methods Enzymol. 2022;670:87-137. doi: 10.1016/bs.mie.2022.03.019. Epub 2022 Apr 23.
Authentic samples of many carotenoids are commercially available, but are often too expensive and of insufficient purity. As an alternative, a wide range of carotenoids can be biosynthesized in recombinant Escherichia coli strains. E. coli cells naturally produce the carotenoid biosynthetic precursors and provide a blank slate for manufacturing different target carotenoids, depending on the introduced genes encoding carotenoid biosynthetic enzymes. This chapter presents a series of plasmids that carry genes encoding carotenoid biosynthetic enzymes and methods to produce in E. coli 21 different carotenoids and two apocarotenoids, which can be used as authentic standards. For the structural confirmation of these carotenoids or apocarotenoids, we show their individual spectral data containing MS/MS and UV-visible, along with their biosynthetic routes in the E. coli transformants.
许多类胡萝卜素的真实样本在商业上是可用的,但通常过于昂贵且纯度不足。作为替代方案,可以在重组大肠杆菌菌株中生物合成广泛的类胡萝卜素。大肠杆菌细胞自然产生类胡萝卜素生物合成前体,并为根据引入的编码类胡萝卜素生物合成酶的基因制造不同的目标类胡萝卜素提供了一个空白模板。本章介绍了一系列携带编码类胡萝卜素生物合成酶的基因的质粒,以及在大肠杆菌中生产 21 种不同类胡萝卜素和两种脱碳类胡萝卜素的方法,这些可以用作真实标准。对于这些类胡萝卜素或脱碳类胡萝卜素的结构确认,我们展示了它们各自的光谱数据,包括 MS/MS 和可见-紫外光谱,以及它们在大肠杆菌转化体中的生物合成途径。