phenylpropanoids

苯丙素类化合物
  • 文章类型: Journal Article
    城市空气污染是一个至关重要的全球性挑战,主要源于城市化和工业活动,不断增加。植被作为空气污染的天然空气过滤器,但是对植物健康的不利影响,光合作用,和新陈代谢可以发生。最近的组学技术彻底改变了分子植物对空气污染反应的研究,克服以前的限制。这篇综述综合了分子植物对主要空气污染物反应的最新进展,强调臭氧(O3),氮氧化物(NOX),和颗粒物(PM)研究。这些污染物诱导其他非生物和生物胁迫常见的应激反应,包括活性氧(ROSs)清除酶和激素信号通路的激活。新的证据表明抗氧化剂酚类化合物生物合成的核心作用,通过苯丙素途径,在空气污染应激反应中。转录因子如WRKY,AP2/ERF,还有MYB,将激素信号与抗氧化剂生物合成联系起来,也受到了影响。迄今为止,研究主要集中在分析单个污染物的实验室研究。这篇综述强调了全面的野外研究和分子耐受性性状鉴定的必要性,这对鉴定耐性植物物种至关重要,旨在开发可持续的基于自然的解决方案(NBS),以减轻城市空气污染。
    Urban air pollution is a crucial global challenge, mainly originating from urbanization and industrial activities, which are continuously increasing. Vegetation serves as a natural air filter for air pollution, but adverse effects on plant health, photosynthesis, and metabolism can occur. Recent omics technologies have revolutionized the study of molecular plant responses to air pollution, overcoming previous limitations. This review synthesizes the latest advancements in molecular plant responses to major air pollutants, emphasizing ozone (O3), nitrogen oxides (NOX), and particulate matter (PM) research. These pollutants induce stress responses common to other abiotic and biotic stresses, including the activation of reactive oxygen species (ROSs)-scavenging enzymes and hormone signaling pathways. New evidence has shown the central role of antioxidant phenolic compound biosynthesis, via the phenylpropanoid pathway, in air pollution stress responses. Transcription factors like WRKY, AP2/ERF, and MYB, which connect hormone signaling to antioxidant biosynthesis, were also affected. To date, research has predominantly focused on laboratory studies analyzing individual pollutants. This review highlights the need for comprehensive field studies and the identification of molecular tolerance traits, which are crucial for the identification of tolerant plant species, aimed at the development of sustainable nature-based solutions (NBSs) to mitigate urban air pollution.
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  • 文章类型: Journal Article
    全球约有三分之一的粮食供应被浪费。酿酒师废谷物(BSG),由啤酒厂大量生产,拥有杰出的营养概况,然而,由于多种原因,它的回收往往被忽视。我们采用整合的宏基因组学和代谢组学技术来评估酶处理和乳酸菌发酵对BSG抗氧化能力的影响。生物处理的BSG显示出改善的抗氧化能力,显著增加(p<0.05)自由基清除活性和类黄酮和多酚含量。非靶向代谢组学显示,乳酸菌发酵导致15种新型抗氧化肽的显著合成(p<0.05),以及显著高于(p<0.05)的异黄酮和苯丙素生物合成途径的富集。相关性分析表明,植物乳杆菌与aucubin和碳水化合物活性酶具有很强的相关性(p<0.05)。即,糖苷水解酶25、糖基转移酶5和糖酯酶9。发酵BSG作为培养基在食品工业中具有潜在的应用,供人类食用的功能性食品成分,和动物的生物活性饲料成分。
    About one-third of the global food supply is wasted. Brewers\' spent grain (BSG), being produced in enormous amounts by the brewery industry, possesses an eminence nutritional profile, yet its recycling is often neglected for multiple reasons. We employed integrated metagenomics and metabolomics techniques to assess the effects of enzyme treatments and Lactobacillus fermentation on the antioxidant capacity of BSG. The biotreated BSG revealed improved antioxidant capability, as evidenced by significantly increased (p < 0.05) radical scavenging activity and flavonoid and polyphenol content. Untargeted metabolomics revealed that Lactobacillus fermentation led to the prominent synthesis (p < 0.05) of 15 novel antioxidant peptides, as well as significantly higher (p < 0.05) enrichment of isoflavonoid and phenylpropanoid biosynthesis pathways. The correlation analysis demonstrated that Lactiplantibacillus plantarum exhibited strong correlation (p < 0.05) with aucubin and carbohydrate-active enzymes, namely, glycoside hydrolases 25, glycosyl transferases 5, and carbohydrate esterases 9. The fermented BSG has potential applications in the food industry as a culture medium, a functional food component for human consumption, and a bioactive feed ingredient for animals.
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  • 文章类型: Journal Article
    干旱限制了菜豆(称为普通豆)的生长和发育。常见的豆类植物含有各种苯丙素类化合物,但尚不清楚这些代谢物的水平是否因干旱而改变。这里,BT6和BT44,两个白豆重组自交系(RIL),在严重干旱下种植。将它们各自的生长和类苯丙烷谱与灌溉良好的植物进行了比较。在严重干旱的情况下,两种RIL在其植被部分积累的生物量要少得多,相对于灌溉良好的植物,它们的根中与更多的phaseolin和phaseollinisophavan相关。在干旱的BT44根中,香豆雌酚的持续积累很明显。在干旱胁迫的BT6和BT44植物中,各种酚酸的叶片轮廓发生了瞬时变化,包括相对于灌溉良好的植物,分别积累了两种单独的咖啡二酸异构体和氯二酸(异构体1)。在干旱胁迫下,BT44中观察到亚铁酸的持续上升,而在BT6中,相对于水分充足的植物,更多的量是短暂的。除了山奈酚二葡萄糖苷(异构体2),大多数叶黄酮苷的浓度没有随干旱而改变。总的来说,在遭受严重干旱的白豆植物中,叶和根苯丙素形态发生了微调。
    Drought limits the growth and development of Phaseolus vulgaris L. (known as common bean). Common bean plants contain various phenylpropanoids, but it is not known whether the levels of these metabolites are altered by drought. Here, BT6 and BT44, two white bean recombinant inbred lines (RILs), were cultivated under severe drought. Their respective growth and phenylpropanoid profiles were compared to those of well-irrigated plants. Both RILs accumulated much less biomass in their vegetative parts with severe drought, which was associated with more phaseollin and phaseollinisoflavan in their roots relative to well-irrigated plants. A sustained accumulation of coumestrol was evident in BT44 roots with drought. Transient alterations in the leaf profiles of various phenolic acids occurred in drought-stressed BT6 and BT44 plants, including the respective accumulation of two separate caftaric acid isomers and coutaric acid (isomer 1) relative to well-irrigated plants. A sustained rise in fertaric acid was observed in BT44 with drought stress, whereas the greater amount relative to well-watered plants was transient in BT6. Apart from kaempferol diglucoside (isomer 2), the concentrations of most leaf flavonol glycosides were not altered with drought. Overall, fine tuning of leaf and root phenylpropanoid profiles occurs in white bean plants subjected to severe drought.
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  • 文章类型: Journal Article
    苯氧基自由基偶联反应在自然界中广泛用于合成复杂分子如木质素。它们在实验室中的使用对于从多酚家族生产高价值化合物具有巨大的潜力。虽然负责产生自由基的酶是众所周知的,后者的行为仍然是神秘的,难以在反应烧瓶中控制。在我们的实验室中使用含有漆酶的B.cinerea酶促分泌组的先前工作表明,二苯乙烯的孵育会导致二聚体,而苯基丙烷类的孵育会导致二聚体以及更大的偶联产物。在这些先前研究的基础上,本文研究了不同结构特征在苯氧基自由基偶联中的作用。我们首先证明了环外共轭双键的存在在产生有效反应中起作用。此外,我们表明,苯丙素类三聚体和四聚体的形成可以通过脱羧反应再生该反应性部分进行。最后,这项研究调查了其他酚类化合物的反应性:二苯乙烯二聚体,一种二氢二苯乙烯,一种4-O-甲基-二苯乙烯和一种具有灰霉病菌酶促分泌组的简单酚。观察到的有效二聚反应始终与共轭到环外双键的对酚的存在相关。缺乏这种结构特征会导致不同的结果,一些化合物显示低转化率或根本没有反应。这项研究允许开发一种可控的方法来合成苯丙素衍生物和新型二苯乙烯衍生物的特定二聚体和四聚体,以及对可以促进有效自由基偶联反应的特征的理解。
    Phenoxy radical coupling reactions are widely used in nature for the synthesis of complex molecules such as lignin. Their use in the laboratory has great potential for the production of high value compounds from the polyphenol family. While the enzymes responsible for the generation of the radicals are well known, the behavior of the latter is still enigmatic and difficult to control in a reaction flask. Previous work in our laboratory using the enzymatic secretome of B. cinerea containing laccases has shown that incubation of stilbenes leads to dimers, while incubation of phenylpropanoids leads to dimers as well as larger coupling products. Building on these previous studies, this paper investigates the role of different structural features in phenoxy radical couplings. We first demonstrate that the presence of an exocyclic conjugated double bond plays a role in the generation of efficient reactions. In addition, we show that the formation of phenylpropanoid trimers and tetramers can proceed via a decarboxylation reaction that regenerates this reactive moiety. Lastly, this study investigates the reactivity of other phenolic compounds: stilbene dimers, a dihydro-stilbene, a 4-O-methyl-stilbene and a simple phenol with the enzymatic secretome of B. cinerea. The observed efficient dimerization reactions consistently correlate with the presence of a para-phenol conjugated to an exocyclic double bond. The absence of this structural feature leads to variable results, with some compounds showing low conversion or no reaction at all. This research has allowed the development of a controlled method for the synthesis of specific dimers and tetramers of phenylpropanoid derivatives and novel stilbene derivatives, as well as an understanding of features that can promote efficient radical coupling reactions.
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  • 文章类型: Journal Article
    癫痫是一种没有明确病因的神经系统疾病,以反复发作的癫痫发作为特征。这些是由于中枢神经系统(CNS)中兴奋性和抑制性神经递质的失调而发生的。精神药物具有不良副作用;许多患者需要一种以上的药物治疗来控制危机。考虑到这一点,这项工作强调从天然产物中发现可以对抗癫痫发作的新物质。使用计算机技术,本综述旨在评估苯丙素衍生物的抗癫痫和多靶点活性。最初,基于配体的虚拟筛选模型(LBVS)与468苯丙素类化合物进行预测生物活性。LBVS是为目标α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)开发的,电压门控钙通道T型(CaV),γ-氨基丁酸A(GABAA),γ-氨基丁酸转运蛋白1型(GAT-1),Q家族的电压门控钾通道(KCNQ),电压门控钠通道(NaV),和N-甲基D-天冬氨酸(NMDA)。分析了在LBVS中具有良好结果的化合物的吸收,分布,新陈代谢,排泄,和毒性(ADMET)参数,后来,在分子对接共识中评估了最佳分子.TR430化合物在药代动力学参数方面表现出最佳结果;其口服吸收率为99.03%,它没有违反任何Lipinski规则,它显示出良好的生物利用度,在评估的参数中,无论是分子还是代谢物都没有观察到细胞毒性。TR430能够与GABAA(激活)和AMPA(抑制)靶标结合,并且表现出良好的结合能和与两个靶标的显著相互作用。所研究的化合物显示是一种有前途的分子,在治疗癫痫的两个基本药理学靶标中具有可能的多靶标活性。
    Epilepsy is a neurological disease with no defined cause, characterized by recurrent epileptic seizures. These occur due to the dysregulation of excitatory and inhibitory neurotransmitters in the central nervous system (CNS). Psychopharmaceuticals have undesirable side effects; many patients require more than one pharmacotherapy to control crises. With this in mind, this work emphasizes the discovery of new substances from natural products that can combat epileptic seizures. Using in silico techniques, this review aims to evaluate the antiepileptic and multi-target activity of phenylpropanoid derivatives. Initially, ligand-based virtual screening models (LBVS) were performed with 468 phenylpropanoid compounds to predict biological activities. The LBVS were developed for the targets alpha- amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), voltage-gated calcium channel Ttype (CaV), gamma-aminobutyric acid A (GABAA), gamma-aminobutyric acid transporter type 1 (GAT-1), voltage-gated potassium channel of the Q family (KCNQ), voltage-gated sodium channel (NaV), and N-methyl D-aspartate (NMDA). The compounds that had good results in the LBVS were analyzed for the absorption, distribution, metabolism, excretion, and toxicity (ADMET) parameters, and later, the best molecules were evaluated in the molecular docking consensus. The TR430 compound showed the best results in pharmacokinetic parameters; its oral absorption was 99.03%, it did not violate any Lipinski rule, it showed good bioavailability, and no cytotoxicity was observed either from the molecule or from the metabolites in the evaluated parameters. TR430 was able to bind with GABAA (activation) and AMPA (inhibition) targets and demonstrated good binding energy and significant interactions with both targets. The studied compound showed to be a promising molecule with a possible multi-target activity in both fundamental pharmacological targets for the treatment of epilepsy.
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  • 文章类型: Journal Article
    水稻(Oryzasativa)是世界近一半人口的主要作物。世界上许多水稻种植地区的地下水中亚砷酸盐和砷酸盐的含量经常升高。同时,与其他主要作物相比,水稻可以积累多达20倍的砷。这使得大量的人面临慢性砷中毒的风险。在这项研究中,根据砷积累引起的生化变化,研究了拉曼光谱(RS)是否可用于诊断水稻中砷的毒性。我们在9天的时间内对水培种植的四个不同水稻品种的亚砷酸盐和砷酸盐胁迫进行了建模。我们的结果表明,从水稻叶片获得的拉曼光谱,结合偏最小二乘判别分析,能够准确检测和识别砷应力,准确率约为89%。我们还对稻叶进行了高效液相色谱(HPLC)分析,以鉴定RS在确认砷中毒中检测到的关键分子分析物。我们发现,由于水稻中亚砷酸盐和砷酸盐的积累,RS主要检测到叶黄素浓度的降低以及香草酸和阿魏酸浓度的增加。这表明这些分子是对砷积累的生化反应的可检测指标。最后,RS与HPLC和ICP-MS的交叉相关性表明RS具有无标记的潜力,非侵入性,和水稻砷积累的无损定量。
    Rice (Oryza sativa) is the primary crop for nearly half of the world\'s population. Groundwater in many rice-growing parts of the world often has elevated levels of arsenite and arsenate. At the same time, rice can accumulate up to 20 times more arsenic compared to other staple crops. This places an enormous amount of people at risk of chronic arsenic poisoning. In this study, we investigated whether Raman spectroscopy (RS) could be used to diagnose arsenic toxicity in rice based on biochemical changes that were induced by arsenic accumulation. We modeled arsenite and arsenate stresses in four different rice cultivars grown in hydroponics over a nine-day window. Our results demonstrate that Raman spectra acquired from rice leaves, coupled with partial least squares-discriminant analysis, enabled accurate detection and identification of arsenic stress with approximately 89% accuracy. We also performed high-performance liquid chromatography (HPLC)-analysis of rice leaves to identify the key molecular analytes sensed by RS in confirming arsenic poisoning. We found that RS primarily detected a decrease in the concentration of lutein and an increase in the concentration of vanillic and ferulic acids due to the accumulation of arsenite and arsenate in rice. This showed that these molecules are detectable indicators of biochemical response to arsenic accumulation. Finally, a cross-correlation of RS with HPLC and ICP-MS demonstrated RS\'s potential for a label-free, non-invasive, and non-destructive quantification of arsenic accumulation in rice.
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  • 文章类型: Journal Article
    国际癌症研究机构(IARC)最近将异丁香酚(2-甲氧基-4-(1-丙烯基)苯酚)归类为可能致癌的人类(2B组)。本研究对常见草药和香料中异丁香酚进行了分析,包括罗勒,肉桂,Ginger,还有肉豆蔻,使用1H核磁共振(NMR)光谱法。此外,通过1H-NMR分析了1300多个咖啡样品的异丁香酚,但在任何分析样本中均未检测到。各种精油,包括肉豆蔻,罗勒,丁香,甜蜜的旗帜,和依兰油,检查异丁香酚含量。在测试的十二种肉豆蔻油中,四个含有异丁香酚,浓度范围为3.68±0.09g/kg至11.2±0.10g/kg。然而,异丁香酚在鱿鱼精油中未检测到,罗勒,依兰,和丁香使用核磁共振光谱。这些发现保证了对以前文献的批判性评估,鉴于一些基质中异丁香酚含量高的报道。毒理学评估已确定,通过肉豆蔻精油接触异丁香酚对人类健康没有风险。
    Isoeugenol (2-methoxy-4-(1-propenyl)phenol) has been recently classified as possibly carcinogenic to humans (Group 2B) by the International Agency for Research on Cancer (IARC). This study conducted an analysis of isoeugenol in common herbs and spices, including basil, cinnamon, ginger, and nutmeg, using 1H nuclear magnetic resonance (NMR) spectrometry. Additionally, over 1300 coffee samples were analysed by 1H-NMR for isoeugenol, but it was not detected in any of the analysed samples. Various essential oils, including nutmeg, basil, clove, sweet flag, and ylang-ylang oils, were examined for isoeugenol content. Out of the twelve nutmeg oils tested, four contained isoeugenol, with concentrations ranging from 3.68 ± 0.09 g/kg to 11.2 ± 0.10 g/kg. However, isoeugenol was not detected in the essential oils of calamus, basil, ylang-ylang, and clove using NMR spectrometry. These findings warrant critical evaluation of the previous literature, given reports of high isoeugenol levels in some of these matrices. A toxicological assessment has determined that there is no risk to human health by exposure to isoeugenol via nutmeg essential oils.
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  • 文章类型: Journal Article
    黄精是一种传统的中药植物,因其在糖尿病和各种慢性疾病中的治疗潜力而备受赞誉。其根茎是富含次生代谢产物的主要功能部位,如类黄酮和皂苷。但是它们的质量因地区而异,对螺旋藻的工业和医药应用提出了挑战。在这项研究中,在清远县和秀水县的球茎中鉴定出482种代谢产物。聚类分析表明,这两个区域之间的样品具有不同的次生代谢物谱。机器学习方法,特别是支持向量机递归特征消除和随机森林,用于进一步鉴定代谢物标记,包括类黄酮,酚酸,和木脂素。进行了比较转录组学和加权基因共表达分析,以发现潜在的候选基因,包括CHI,UGT1和PcOMT10/11/12/13与这些化合物相关。使用烟草瞬时表达系统的功能测定表明,PcOMT10/11/12/13确实影响了类苯丙烷途径和类苯丙烷相关代谢物的代谢通量,例如β-6,8-二-C-葡萄糖苷,syringaresinol-4'-O-吡喃葡萄糖苷,和1-O-西洋基-D-葡萄糖。这些发现确定了这两个区域之间的代谢物标记,并为工程这些化合物的生物合成提供了有价值的遗传见解。
    Polygonatum cyrtonema Hua is a traditional Chinese medicinal plant acclaimed for its therapeutic potential in diabetes and various chronic diseases. Its rhizomes are the main functional parts rich in secondary metabolites, such as flavonoids and saponins. But their quality varies by region, posing challenges for industrial and medicinal application of P. cyrtonema. In this study, 482 metabolites were identified in P. cyrtonema rhizome from Qingyuan and Xiushui counties. Cluster analysis showed that samples between these two regions had distinct secondary metabolite profiles. Machine learning methods, specifically support vector machine-recursive feature elimination and random forest, were utilized to further identify metabolite markers including flavonoids, phenolic acids, and lignans. Comparative transcriptomics and weighted gene co-expression analysis were performed to uncover potential candidate genes including CHI, UGT1, and PcOMT10/11/12/13 associated with these compounds. Functional assays using tobacco transient expression system revealed that PcOMT10/11/12/13 indeed impacted metabolic fluxes of the phenylpropanoid pathway and phenylpropanoid-related metabolites such as chrysoeriol-6,8-di-C-glucoside, syringaresinol-4\'-O-glucopyranosid, and 1-O-Sinapoyl-D-glucose. These findings identified metabolite markers between these two regions and provided valuable genetic insights for engineering the biosynthesis of these compounds.
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  • 文章类型: Journal Article
    褪黑激素,多效性小分子,用于园艺作物以延缓衰老并保持采后质量。在这项研究中,100µM褪黑素治疗延迟了色差指数h*和a*的下降,保持叶绿素和类胡萝卜素的含量,从而延缓了羽衣甘蓝的黄化和衰老。转录组分析明确验证了褪黑激素在20°C贮藏的羽衣甘蓝中延缓叶片衰老的功效。经过三天的储存期,与对照组相比,褪黑素治疗组表现出1637个差异表达基因(DEGs).DEG分析阐明褪黑激素诱导的抗衰老主要控制苯丙素生物合成,脂质代谢,植物信号转导,和钙信号转导。褪黑素处理上调与一般苯丙素生物合成相关的核心酶基因,类黄酮生物合成,和α-亚麻酸生物合成途径。它影响了木质素代谢通量的重定向,抑制茉莉酸和脱落酸信号转导,同时刺激生长素信号转导。此外,褪黑素治疗下调RBOH表达和上调编码CaM的基因,从而影响钙信号转导。这项研究强调了褪黑激素是延缓叶片衰老的有希望的方法,并提供了对采后中国羽衣甘蓝中褪黑激素介导的抗衰老机制的见解。
    Melatonin, a pleiotropic small molecule, is employed in horticultural crops to delay senescence and preserve postharvest quality. In this study, 100 µM melatonin treatment delayed a decline in the color difference index h* and a*, maintaining the content of chlorophyll and carotenoids, thereby delaying the yellowing and senescence of Chinese kale. Transcriptome analysis unequivocally validates melatonin\'s efficacy in delaying leaf senescence in postharvest Chinese kale stored at 20 °C. Following a three-day storage period, the melatonin treatment group exhibited 1637 differentially expressed genes (DEGs) compared to the control group. DEG analysis elucidated that melatonin-induced antisenescence primarily governs phenylpropanoid biosynthesis, lipid metabolism, plant signal transduction, and calcium signal transduction. Melatonin treatment up-regulated core enzyme genes associated with general phenylpropanoid biosynthesis, flavonoid biosynthesis, and the α-linolenic acid biosynthesis pathway. It influenced the redirection of lignin metabolic flux, suppressed jasmonic acid and abscisic acid signal transduction, and concurrently stimulated auxin signal transduction. Additionally, melatonin treatment down-regulated RBOH expression and up-regulated genes encoding CaM, thereby influencing calcium signal transduction. This study underscores melatonin as a promising approach for delaying leaf senescence and provides insights into the mechanism of melatonin-mediated antisenescence in postharvest Chinese kale.
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  • 文章类型: Journal Article
    背景:苯丙素类是一类具有多种生物学功能的植物次生代谢产物,源自芳香氨基酸。蓝细菌是可持续生产植物类苯丙素的有前途的宿主生物。我们以前设计了集胞藻。PCC6803生产反式肉桂酸(tCA)和对香豆酸(pCou),苯丙素途径的第一个中间体,通过过表达苯丙氨酸和酪氨酸氨裂解酶。在这项研究中,我们的目的是提高集胞藻中目标化合物tCA和pCou的产量。
    结果:我们消除了4-羟基苯基丙酮酸双加氧酶(HPPD)活性,这是一个消耗酪氨酸的竞争途径,可能,用于生育酚合成的苯丙氨酸。此外,莽草酸途径末端步骤的几个基因单独或在操纵子中过表达,如芳香转氨酶,来自运动发酵单胞菌的反馈不敏感的环己二烯基脱氢酶(TyrC)和来自大肠杆菌的融合的分支酸变位酶/预苯酸脱水酶的分支酸变位酶(CM)结构域。与亲本生产菌株相比,敲除HPPD后,获得的工程菌株的tCA和pCou产量提高了近1.5倍,光合自养生长7天后,积累了138±3.5mgL-1的tCA和72.3±10.3mgL-1的pCou。然而,当任何通路基因过表达时,没有进一步的改善.最后,我们使用先前获得的AtPRM8和TsPRM8集胞藻菌株,这些菌株具有shikimate途径的失调,作为过表达具有ppd敲除的合成构建体的背景。
    结论:消除HPPD可在相似程度上提高tCA和pCou生产力。使用基于PRM8的菌株作为过表达合成构建体的背景,然而,没有提高tCA和pCou滴度,这表明对苯丙氨酸和酪氨酸合成的末端步骤进行了严格的调节。这项工作有助于建立蓝细菌作为苯丙烷生产的宿主。
    BACKGROUND: Phenylpropanoids are a large group of plant secondary metabolites with various biological functions, derived from aromatic amino acids. Cyanobacteria are promising host organisms for sustainable production of plant phenylpropanoids. We have previously engineered Synechocystis sp. PCC 6803 to produce trans-cinnamic acid (tCA) and p-coumaric acid (pCou), the first intermediates of phenylpropanoid pathway, by overexpression of phenylalanine- and tyrosine ammonia lyases. In this study, we aimed to enhance the production of the target compounds tCA and pCou in Synechocystis.
    RESULTS: We eliminated the 4-hydroxyphenylpyruvate dioxygenase (HPPD) activity, which is a competing pathway consuming tyrosine and, possibly, phenylalanine for tocopherol synthesis. Moreover, several genes of the terminal steps of the shikimate pathway were overexpressed alone or in operons, such as aromatic transaminases, feedback insensitive cyclohexadienyl dehydrogenase (TyrC) from Zymomonas mobilis and the chorismate mutase (CM) domain of the fused chorismate mutase/prephenate dehydratase enzyme from Escherichia coli. The obtained engineered strains demonstrated nearly 1.5 times enhanced tCA and pCou production when HPPD was knocked out compared to the parental production strains, accumulating 138 ± 3.5 mg L-1 of tCA and 72.3 ± 10.3 mg L-1 of pCou after seven days of photoautotrophic growth. However, there was no further improvement when any of the pathway genes were overexpressed. Finally, we used previously obtained AtPRM8 and TsPRM8 Synechocystis strains with deregulated shikimate pathway as a background for the overexpression of synthetic constructs with ppd knockout.
    CONCLUSIONS: HPPD elimination enhances the tCA and pCou productivity to a similar extent. The use of PRM8 based strains as a background for overexpression of synthetic constructs, however, did not promote tCA and pCou titers, which indicates a tight regulation of the terminal steps of phenylalanine and tyrosine synthesis. This work contributes to establishing cyanobacteria as hosts for phenylpropanoid production.
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