Ritsema Tita, Joling Jeanine, Smeekens Sjef
Department of Molecular Plant Physiology, Utrecht University, H.R. Kruytgebouw, Padualaan 8, NL-3584 CH Utrecht, The Netherlands.
New Phytol. 2003 Oct;160(1):61-67. doi: 10.1046/j.1469-8137.2003.00858.x.
• Fructan : fructan 6G-fructosyltransferase (6G-FFT) has been proposed to be the enzyme essential for the production of neo-series inulin. Transfer of a fructose unit from short chain inulins to the C6 of the glucose residue of sucrose or inulin was proposed to be its most important characteristic. Here, we investigate the activity of 6G-FFT from onion (Allium cepa) more thoroughly. • Tobacco BY2 suspension cultures were employed as an expression system for the fructosyltransferase 6G-FFT. Activity was measured using 1-kestose as a substrate and products were detected using high-performance anion exchange chromatography (HPAEC). • 6G-FFT showed multiple activities. An array of fructans of the inulin series and inulin neo-series were produced. First 1,1-kestotetraose and 1 and 6G-kestotetraose were synthesized, as well as 6G-kestotriose. Prolonged incubation produces a complex fructan series with a higher degree of polymerization. • The fructan pattern observed after incubation of onion 6G-FFT with 1-kestose closely resembles the complex fructan pattern found in onion. These results questions the need for a separate fructan : fructan 1-fructosyltransferase (1-FFT) activity in onion.
• 果聚糖:果聚糖6G-果糖基转移酶(6G-FFT)被认为是新系列菊粉产生所必需的酶。短链菊粉中的果糖单元转移至蔗糖或菊粉葡萄糖残基的C6被认为是其最重要的特征。在此,我们更深入地研究了洋葱(葱属)中6G-FFT的活性。
• 烟草BY2悬浮培养物被用作果糖基转移酶6G-FFT的表达系统。以1-蔗果三糖为底物测定活性,并使用高效阴离子交换色谱法(HPAEC)检测产物。
• 6G-FFT表现出多种活性。产生了一系列菊粉系列和菊粉新系列的果聚糖。首先合成了1,1-蔗果四糖、1和6G-蔗果四糖以及6G-蔗果三糖。长时间孵育会产生具有更高聚合度的复杂果聚糖系列。
• 洋葱6G-FFT与1-蔗果三糖孵育后观察到的果聚糖模式与洋葱中发现的复杂果聚糖模式非常相似。这些结果质疑了洋葱中是否需要单独的果聚糖:果聚糖1-果糖基转移酶(1-FFT)活性。