Tian Bing, Sun Zongtao, Xu Zhenjian, Shen Shaochuan, Wang Hu, Hua Yuejin
Key Laboratory for Nuclear-Agricultural Sciences of Chinese Ministry of Agriculture and Zhejiang Province, Institute of Nuclear-Agricultural Sciences, Zhejiang University, 310029 Hangzhou, PR China.
Microbiology (Reading). 2008 Dec;154(Pt 12):3697-3706. doi: 10.1099/mic.0.2008/021071-0.
Deinococcus radiodurans strain R1 synthesizes deinoxanthin, a unique carotenoid product, which contributes to cell resistance following various stresses. The biosynthetic pathway of deinoxanthin is unclear, although several enzymes are presumed to be involved. The gene (dr2250) predicted by gene homologue analysis to encode carotenoid 3',4'-desaturase (CrtD) was deleted to investigate its function. A mutant deficient in the gene homologue of crtLm (dr0801) was also constructed to verify the catalytic function of the gene product in the native host. Carotenoid analysis of the resultant mutants verified that DR2250 encodes carotenoid 3',4'-desaturase, which catalyses the C-3',4'-desaturation of the monocyclic precursor of deinoxanthin but not acyclic carotenoids. Mutation of the gene homologue of crtLm (dr0801) resulted in accumulation of lycopene, confirming that it encodes the lycopene cyclase in the native host. The lack of CrtD decreased the antioxidant capacity of the mutant deficient in dr2250 compared with the wild-type, indicating that the C-3',4'-desaturation step contributes to the antioxidant capacity of deinoxanthin in D. radiodurans.
耐辐射球菌R1菌株合成脱氮叶黄素,一种独特的类胡萝卜素产物,它有助于细胞在各种应激后产生抗性。尽管推测有几种酶参与其中,但脱氮叶黄素的生物合成途径尚不清楚。通过基因同源性分析预测编码类胡萝卜素3',4'-去饱和酶(CrtD)的基因(dr2250)被删除以研究其功能。还构建了crtLm基因同源物(dr0801)缺陷的突变体,以验证该基因产物在天然宿主中的催化功能。对所得突变体的类胡萝卜素分析证实,DR2250编码类胡萝卜素3',4'-去饱和酶,它催化脱氮叶黄素单环前体的C-3',4'-去饱和反应,但不催化无环类胡萝卜素的反应。crtLm基因同源物(dr0801)的突变导致番茄红素积累,证实它在天然宿主中编码番茄红素环化酶。与野生型相比,缺乏CrtD降低了dr2250缺陷突变体的抗氧化能力,表明C-3',4'-去饱和步骤有助于耐辐射球菌中脱氮叶黄素的抗氧化能力。