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首页> 外文学位 >Biosynthesis of p-hydroxyphenyl-2-butanone and its correlation to primary metabolism in cell suspension cultures of Rubus idaeus.
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Biosynthesis of p-hydroxyphenyl-2-butanone and its correlation to primary metabolism in cell suspension cultures of Rubus idaeus.

机译：对羟基苯-2-丁酮的生物合成及其与悬钩子悬液培养中初级代谢的关系。

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The principle objective of this research was to investigate carbon substrate metabolism in the biosynthesis of p-hydroxyphenyl-2-butanone (p-HPB2) in Rubus idaeus (raspberry). This was achieved by feeding various precursors and through detailed stoichiometric modeling of the biosynthetic pathway. The typical cell culture profiles for the flavor component (p-HPB2), its intermediate precursor p-hydroxyphenylbut-3-ene-2-one (p-HPB 1) and a distant precursor p-coumaric acid (p-CA) were measured and related to carbohydrate consumption and cell proliferation. The transient production of these metabolites and lack of sequential changes in concentration along the pathway indicated a complex regulatory process controlling their production. The signal coupler methyl jasmonate was found to induce the synthesis of p-HPB2 and p-HPB 1. Its effect was a function of elicitor concentration, exposure time and time of addition.; The link between primary and secondary metabolism was investigated by metabolic modeling and by feeding precursors involved in p-HPB 2 formation. Exogenous addition of precursors (sucrose, L-phenylalanine, p-CA and p-HPB 1) indicated a possible limitation in the phenylpropanoid pathway. Sucrose fed cultures exhibited negligible change in sucrose consumption rates from control cultures, thus implying a self-regulatory step in sucrose uptake and further, that sucrose is not a limiting substrate for p-HPB2 synthesis. Lack of p-CA increase in sucrose fed cultures, suggested a preferential channeling of the fed sucrose via the glycolysis into p-HPB2 formation. A synergistic effect of combination feeds on metabolite levels demonstrated the requirement of primary and secondary metabolites for p-HPB2 synthesis. The rapid utilization of fed precursors without any intracellular accumulation established that they are not stable end products. Modeling the biosynthetic pathway via metabolic flux analysis was carried out to generate a stoichiometric equation that indicates a maximum theoretical yield of p-HPB2 from sucrose of 0.46 mol/mol. Metabolic modeling further demonstrated that majority of the carbon flux (60%–70%) is channelled via glycolysis, thus confirming the experimental observation of preferential channeling of carbon substrate.
机译：这项研究的主要目的是研究 p -羟基苯基-2-丁酮（ p -HPB 2 ）生物合成中的碳底物代谢。在 Rubus idaeus （覆盆子）中。这是通过进料各种前体并通过生物合成途径的详细化学计量模型实现的。风味成分（ p -HPB 2 ）及其中间前体 p -羟基苯基丁-3-烯-2-的典型细胞培养谱测量了一种（ p -HPB 1 ）和远距离的前体对香豆酸（ p -CA），它们与碳水化合物的消耗和细胞相关增殖。这些代谢物的瞬时产生和沿该路径浓度的连续变化均未表明控制其产生的复杂调控过程。发现茉莉酸甲酯信号耦合剂诱导 p -HPB 2 和 p -HPB 1 的合成。其作用是激发子浓度，暴露时间和添加时间的函数。通过代谢模型和喂养参与 p -HPB 2 形成的前体来研究初次和二次代谢之间的联系。外源添加前体（蔗糖， L -苯丙氨酸， p -CA和 p -HPB 1 ）表明苯丙烷途径的可能限制。饲喂蔗糖的培养物与对照培养物相比，蔗糖消耗率的变化可忽略不计，因此暗示了蔗糖摄取的自我调节步骤，此外，蔗糖不是 p -HPB 2 < / sub>综合。蔗糖喂养的培养物中缺乏 p -CA的增加，这表明通过糖酵解将饲喂的蔗糖优先引导形成 p -HPB 2 通道。复合饲料对代谢物水平的协同作用表明 p -HPB 2 合成需要一级和二级代谢产物。饲料前体的快速利用没有任何细胞内积累，证明它们不是稳定的最终产品。通过代谢通量分析对生物合成途径进行建模，以生成化学计量方程，该方程表明蔗糖的 p -HPB 2 的最大理论产率为0.46 mol / mol。代谢建模进一步证明，大部分碳通量（60％–70％）是通过糖酵解引导的，从而证实了对碳底物优先引导的实验观察。

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