Schofield Andrew, Paliyath Gopinadhan
Department of Food Science, University of Guelph, Guelph, Ont., N1G 2W1 Canada.
Plant Physiol Biochem. 2005 Dec;43(12):1052-60. doi: 10.1016/j.plaphy.2005.10.006. Epub 2005 Dec 13.
The accumulation of carotenoids greatly influences the quality of ripe tomato (Solanum lycopersicum) fruit because these compounds contribute to their color and functional food value. The regulatory properties and gene expression of phytoene synthase (PSY), a key enzyme involved in carotenoid biosynthesis, was analyzed in tomato fruit. PSY activity was detected in the soluble fraction showing a time- and a protein-dependent increase in the formation of phytoene from geranylgeranyl pyrophosphate (GGPP). The enzyme was stimulated by Mg(2+) and ATP, and was inhibited by Tween 20 and Triton X-100. The enzyme followed typical Michaelis-Menten kinetics with increasing GGPP. During in vivo studies, pericarp discs from breaker stage tomatoes were ripened in darkness (D), or D interrupted by daily pulses of red (R) light, or R followed by far red (FR) light. After 14 days incubation, R-treated discs had accumulated 12 mg carotenoids/100 g fresh weight; nearly a 50% increase over D- and R/FR-treated discs. This R/FR reversibility of carotenoid accumulation was also observed in PSY activity 8 days post breaker, where it showed peak activity. However, this R/FR regulation of PSY activity was not reflected in PSY1 transcript levels, suggesting that PSY may be subject to post-translational regulation.
类胡萝卜素的积累极大地影响成熟番茄(Solanum lycopersicum)果实的品质,因为这些化合物决定了果实的颜色和功能性食品价值。本研究分析了参与类胡萝卜素生物合成的关键酶——八氢番茄红素合酶(PSY)的调控特性和基因表达。在可溶性部分检测到PSY活性,表明其以时间和蛋白质依赖性方式从牻牛儿基牻牛儿基焦磷酸(GGPP)形成八氢番茄红素。该酶受到Mg(2+)和ATP的刺激,并被吐温20和曲拉通X-100抑制。随着GGPP浓度增加,该酶遵循典型的米氏动力学。在体内研究中,将绿熟期番茄的果皮切片在黑暗(D)中成熟,或在黑暗中每天用红光(R)脉冲打断,或先用红光再用远红光(FR)处理。孵育14天后,用红光处理的切片积累了12mg类胡萝卜素/100g鲜重;比黑暗处理和红光/远红光处理的切片增加了近50%。在破色期后8天的PSY活性中也观察到类胡萝卜素积累的这种红光/远红光可逆性,此时PSY活性达到峰值。然而,PSY活性的这种红光/远红光调节并未反映在PSY1转录水平上,这表明PSY可能受到翻译后调控。