van der Hoeven R S, Steffens J C
Department of Plant Breeding and Biometry, 252 Emerson Hall, Cornell University, Ithaca, New York 14853, USA.
Plant Physiol. 2000 Jan;122(1):275-82. doi: 10.1104/pp.122.1.275.
Short- and medium-chain-length fatty acids (FAs) are important constituents of a wide array of natural products. Branched and straight short-chain-length FAs originate from branched chain amino acid metabolism, and serve as primers for elongation in FA synthase-like reactions. However, a recent model proposes that the one-carbon extension reactions that utilize 2-oxo-3-methylbutyric acid in leucine biosynthesis also catalyze a repetitive one-carbon elongation of short-chain primers to medium-chain-length FAs. The existence of such a mechanism would require a novel form of regulation to control carbon flux between amino acid and FA biosynthesis. A critical re-analysis of the data used to support this pathway fails to support the hypothesis for FA elongation by one-carbon extension cycles of alpha-ketoacids. Therefore, we tested the hypothesis experimentally using criteria that distinguish between one- and two-carbon elongation mechanisms: (a) isotopomer patterns in terminal carbon atom pairs of branched and straight FAs resulting from differential labeling with [(13)C]ăcetate; (b)(13)C]threonine labeling patterns in odd- and even chain length FAs; and (c) differential sensitivity of elongation reactions to inhibition by cerulenin. All three criteria indicated that biosynthesis of medium-chain length FAs is mediated primarily by FA synthase-like reactions.
短链和中链脂肪酸(FAs)是多种天然产物的重要组成部分。支链和直链短链脂肪酸源自支链氨基酸代谢,并在类脂肪酸合酶反应中作为延伸的引物。然而,最近的一个模型提出,在亮氨酸生物合成中利用2-氧代-3-甲基丁酸的一碳延伸反应也催化短链引物向中链脂肪酸的重复一碳延伸。这种机制的存在将需要一种新的调节形式来控制氨基酸和脂肪酸生物合成之间的碳通量。对用于支持该途径的数据进行的关键重新分析未能支持α-酮酸的一碳延伸循环导致脂肪酸延伸的假设。因此,我们使用区分一碳和二碳延伸机制的标准对该假设进行了实验测试:(a)用[(13)C]醋酸盐进行差异标记后,支链和直链脂肪酸末端碳原子对中的同位素异构体模式;(b)奇数和偶数链长脂肪酸中的[(13)C]苏氨酸标记模式;以及(c)延伸反应对浅蓝菌素抑制的不同敏感性。所有这三个标准都表明,中链脂肪酸的生物合成主要由类脂肪酸合酶反应介导。