Henke Nadja A, Heider Sabine A E, Hannibal Silvin, Wendisch Volker F, Peters-Wendisch Petra
Genetics of Prokaryotes, Faculty of Biology, Center for Biotechnology, Bielefeld UniversityBielefeld, Germany.
Front Microbiol. 2017 Apr 24;8:633. doi: 10.3389/fmicb.2017.00633. eCollection 2017.
is a natural producer of the C50 carotenoid decaprenoxanthin. The operon comprises most of its genes for terpenoid biosynthesis. The MarR-type regulator encoded upstream and in divergent orientation of the carotenoid biosynthesis operon has not yet been characterized. This regulator, named CrtR in this study, is encoded in many actinobacterial genomes co-occurring with terpenoid biosynthesis genes. CrtR was shown to repress the operon of since DNA microarray experiments revealed that transcript levels of operon genes were increased 10 to 70-fold in its absence. Transcriptional fusions of a promoter-less gene with the operon and promoters confirmed that CrtR represses its own gene and the operon. Gel mobility shift assays with purified His-tagged CrtR showed that CrtR binds to a region overlapping with the -10 and -35 promoter sequences of the operon. Isoprenoid pyrophosphates interfered with binding of CrtR to its target DNA, a so far unknown mechanism for regulation of carotenogenesis. The molecular details of protein-ligand interactions remain to be studied. Decaprenoxanthin synthesis by wild type was enhanced 10 to 30-fold upon deletion of and was decreased 5 to 6-fold as result of overexpression. Moreover, deletion of was shown as metabolic engineering strategy to improve production of native and non-native carotenoids including lycopene, β-carotene, C.p. 450 and sarcinaxanthin.
是C50类胡萝卜素癸异戊烯基叶黄素的天然生产者。该操纵子包含其大部分萜类生物合成基因。在类胡萝卜素生物合成操纵子上游以反向方向编码的MarR型调节因子尚未得到表征。本研究中命名为CrtR的这种调节因子,在许多与萜类生物合成基因同时出现的放线菌基因组中编码。由于DNA微阵列实验表明在其缺失时操纵子基因的转录水平增加了10至70倍,因此CrtR被证明可抑制该操纵子。一个无启动子基因与操纵子和启动子的转录融合证实CrtR抑制其自身基因和操纵子。用纯化的His标签CrtR进行的凝胶迁移率变动分析表明,CrtR与操纵子-10和-35启动子序列重叠的区域结合。类异戊二烯焦磷酸干扰CrtR与其靶DNA的结合,这是一种迄今为止未知的类胡萝卜素生成调节机制。蛋白质-配体相互作用的分子细节仍有待研究。野生型的癸异戊烯基叶黄素合成在缺失后增强了10至30倍,而在过表达后降低了5至6倍。此外,缺失被证明是一种代谢工程策略,可提高包括番茄红素、β-胡萝卜素、C.p. 450和肌动球菌黄素在内的天然和非天然类胡萝卜素的产量。