葡萄酒苹果酸-乳酸菌精氨酸代谢研究概况

葡萄酒苹果酸-乳酸菌的精氨酸代谢会导致葡萄酒中氨基甲酸乙酯含量的增加,从而严重影响葡萄酒的饮用安全性。近年来研究表明,葡萄酒苹果酸-乳酸菌的精氨酸代谢途径是精氨酸脱亚氨基酶途径(Arginine deiminasepathway,简称ADI途径)。系统分析苹果酸-乳酸菌的ADI途径、精氨酸转运机制、ADI途径酶的调节等方面的研究进展,阐明葡萄酒苹果酸-乳酸菌的精氨酸代谢对酿造优质葡萄酒具有重要的理论和实际意义。     

植物精氨酸及其代谢产物的生理功能

L-精氨酸在植物中除作为一种重要的氮素贮藏营养物供再利用外,还是生成多胺（PA）和-氧化氮（NO）等的前体物质,而PA和NO都是植物中重要的信使分子,参与包括生长发育、抗逆性等在内的几乎所有的生理生化过程。精氨酸脱羧酶（ADC）、精氨酸酶和一氧化氮合酶（NOS）是L-精氨酸分解代谢的关键酶,精氨酸可经ADC或精氨酸酶-鸟氨酸脱羧酶（ODC）途径形成PA,也可经NOS途径形成NO,3个酶活性的相对强弱,决定了精氨酸的代谢方向。根系在越冬期间会积累丰富的精氨酸;精氨酸代谢对于植物感知和适应环境变化有重要意义。     

L-精氨酸生物合成机制及其代谢工程育种研究进展

L-精氨酸是人体半必需的氨基酸,在生命代谢过程中起着非常重要的作用,且具有广泛的应用价值及市场需求。目前,L-精氨酸主要采用微生物发酵法进行生产,为了提高L-精氨酸的产量和稳定性,最有效的方法是优化L-精氨酸生产菌株,通过代谢工程改造微生物菌株有望达到这一目的。本文分析了微生物中L-精氨酸的代谢途径和调控机制,并综述了构建高产L-精氨酸的代谢工程策略。此外,展望了菌株稳定性和底物扩展利用的未来研究方向。     

内源性代谢分子——精氨酸/一氧化氮调节机体生理功能北大核心CSCD

精氨酸是人体中功能最多的氨基酸,作为多种内源性代谢产物如多胺、鸟氨酸、一氧化氮(nitric oxide,NO)等的前体物质,它在人体正常稳态的调节中中具有重要的生理功能。其中NO通过其特殊的理化性质及代谢过程在人体各个系统中担当着要角色。NO自被发现以来一直活跃在生命科学的前沿领域。但直到目前,NO的生理及病理作用仍然有许多问题有待更加深入地研究。本文对精氨酸/NO代谢途径及其中间代谢产物对机体正常生理功能、自稳态调节做一个简要的综述。     

精氨酸抗氧化作用机制

精氨酸是一种功能性氨基酸,在机体生理功能、新陈代谢和营养等方面发挥着重要作用。精氨酸具有抗氧化能力。目前的体外研究表明精氨酸具有较强的清除DPPH自由基、ABTS自由基、超氧自由基能力以及一定的还原力。作为一种带电子的碱性氨基酸,精氨酸可能通过胍基基团向自由基提供电子并与其作用,终止自由基链式反应,从而显示出还原能力与体外抗氧化能力。体内实验则表明精氨酸能有效地提高机体总抗氧化能力,降低体内自由基含量,抑制ROS生成与积累,促进谷胱甘肽(GSH)合成与积累,增强内源性抗氧化酶(CAT、SOD、GPx等)活性,抑制氧化应激的产生。精氨酸能够通过精氨酸——一氧化氮途径、GSH途径、Nrf2信号通路途径及其他途径发挥体内抗氧化作用。本文主要综述了目前精氨酸体外与体内抗氧化功能及其相关作用机制的研究进展,为精氨酸的实际应用提供理论指导意义。     

L-精氨酸合成及高产菌选育与发酵研究进展

精氨酸是合成蛋白质的重要原料,是一些代谢途径的中间代谢物.它在人和动物体内具有重要的生理生化功能,在食品与医药工业应用十分广泛.对精氨酸高产菌株选育、发酵工艺优化、真核生物中的合成途径、代谢调控机制等方面最新研究进展做了综述.     

嗜盐四联球菌arc operon多拷贝基因在精氨酸代谢中的作用

【背景】嗜盐四联球菌(Tetragenococcus halophilus)是一类存在于发酵食品中的耐盐乳酸菌,研究其精氨酸(arginine,Arg)代谢对解析食品发酵过程中氨基甲酸乙酯(ethyl carbamate,EC)前体积累机制和保障食品安全具有重要意义。【目的】研究酱醪来源嗜盐四联球菌精氨酸脱亚氨基(arginine deiminase,ADI)途径的基因构成,揭示这些基因对菌株精氨酸代谢和氨基甲酸乙酯前体瓜氨酸(citrulline,Cit)利用与积累的影响。【方法】采用PCR扩增与测序分析不同菌株的ADI途径基因组成,通过比较ADI途径关键基因转录水平和关键酶活性,探究环境因素对嗜盐四联球菌代谢氨基酸能力的影响及各拷贝基因参与氨基酸代谢的功能。【结果】酱醪来源嗜盐四联球菌基因组中ADI途径基因类型主要有两大类：以菌株R23为代表含有完整arc操纵子(operon)基因且具有最多基因拷贝数;以菌株C3为代表缺失arcA和arcB但含有多拷贝arcB和arcC。基因组中有arcA的菌株才具有利用精氨酸能力,并通过利用精氨酸生成瓜氨酸。体系中精氨酸含量和乙醇与脂肪酸的存在均可影响嗜盐四联球菌利用精氨酸积累中间产物瓜氨酸。当精氨酸含量大于5 g/L或体系中含有乙醇与脂肪酸时,嗜盐四联球菌会利用精氨酸积累中间产物瓜氨酸。脂肪酸和乙醇对ADI途径的3个关键酶均有显著抑制作用,可使精氨酸脱亚氨基酶(arginine deiminase,ADI)、鸟氨酸氨甲酰基转移酶(ornithine transcarbamylase,OTC)和氨甲酰磷酸激酶(carbamate kinase,CK)的活性分别降低41.0%、46.4%和60.0%。嗜盐四联球菌中arcB转录水平分别是其拷贝arcB₁和arcB₂的10.5倍和29.8倍,arcC的转录水平分别是arcC₁、arcC₂、arcC₃的17.6、20.3、23.9倍,说明arcB和arcC在瓜氨酸代谢中起主要作用。【结论】精氨酸含量和乙醇加脂肪酸是影响嗜盐四联球菌代谢精氨酸能否积累瓜氨酸的关键环境因素。嗜盐四联球菌arc operon的多拷贝基因中,arcB和arcC基因在瓜氨酸代谢中起主要作用。     

精氨酸代谢途径抗酸关键基因对乳酸乳球菌Lactococcus lactis NZ9000胁迫抗性的影响

【目的】寻找精氨酸代谢途径中与酸胁迫相关的关键作用因素。【方法】通过在Lactococcus lactis NZ9000中分别过量表达来源于Lactobacillus casei Zhang的精氨酰琥珀酸合成酶(ASS)和精氨酰琥珀酸裂解酶(ASL)改变精氨酸代谢提高酸胁迫抗性。【结果】与对照菌株对比,重组菌株在环境胁迫下表现了较高的生长性能、存活率和发酵性能。生理学分析发现,酸胁迫环境下,重组菌株细胞有较高的胞内NH4+、ATP含量和H+-ATPase活性,并显著提高了精氨酸脱亚胺酶(ADI)途径中的氨基酸浓度。进一步的转录分析发现,天冬氨酸合成、精氨酸代谢相关的基因转录水平上调。【结论】在L.lactis NZ9000中过量表达ASS或ASL可以引发精氨酸代谢流量的上调,进而提高了细胞的多种胁迫抗性。精氨酸合成途径广泛存在于多种微生物中,为微生物,尤其是工业微生物提高胁迫抗性提供了新思路。     

原核细胞精氨酸生物合成途径的研究进展

原核生物的精氨酸生物合成包含8个酶系,起始于乙酰谷氨酸激酶催化的谷氨酸的乙酰化。到第五步乙酰基团脱离,乙酰谷氨酸通过3个酶的作用,进一步合成乙酰化中间产物。鸟氨酸被氨甲酰基化生成瓜氨酸,天冬氨酸介入后形成精氨琥珀酸,最后形成终产物精氨酸。主要就精氨酸生物合成途径、合成过程中主要酶系及反馈抑制蛋白的作用机制进行了概述。此外,提出了目前精氨酸代谢研究中存在的问题及未来的研究方向。     

精氨酸及其代谢产物对缺血性脑卒中的作用

摘要：缺血性脑卒中是成年人群致残、致死的重要原因之一,有效治疗手段和药物的匮乏是脑卒中致残的主要原因。精氨酸既是一种营养物质,又具有多种独特的生理与药理作用,在早产儿和严重应激状态下精氨酸在维持正氮平衡与正常生理功能方面发挥重要作用,常将精氨酸称为条件必需氨基酸。精氨酸是生物体合成多胺的前体物质,同时精氨酸代谢也产生高活性自由基一氧化氮。精氨酸代谢及其代谢产物的改变可对脑卒中产生多种影响,如线粒体功能破坏、钙离子通道紊乱、血脑屏障损伤等。本文综述了精氨酸及其代谢产物在缺血性脑卒中病理过程中的作用。深入的研究和探讨其损伤和保护的双重作用机制将为缺血性脑卒中的防御和治疗提供新的策略。     

Agmatine and Imidazoline Receptors: Their Role in Opioid Analgesia, Tolerance and Dependence

Agmatine is an endogenous amine that is synthesized following the decarboxylation of L-arginine by arginine decarboxylase. Agmatine exists in mammalian brain and has been proposed as a neurotransmitter and/or neurotransmodulator. Agmatine binds to several targets and is considered as an endogenous ligand for imidazoline receptors. This review, mainly based on our research work in the past decade, focused on the modulations by agmatine action on imidazoline receptors to opioid analgesia, tolerance and dependence, and its possible neurochemical mechanisms. We went on to propose that agmatine and imidazoline receptors constitute a novel system of modulating opioid functions.     

Arabidopsis polyamine biosynthesis: absence of ornithine decarboxylase and the mechanism of arginine decarboxylase activity

Unlike other eukaryotes, which can synthesize polyamines only from ornithine, plants possess an additional pathway from arginine. Occasionally non-enzymatic decarboxylation of ornithine could be detected in Arabidopsis extracts; however, we could not detect ornithine decarboxylase (ODC; EC 4. 1.1.17) enzymatic activity or any activity inhibitory to the ODC assay. There are no intact or degraded ODC sequences in the Arabidopsis genome and no ODC expressed sequence tags. Arabidopsis is therefore the only plant and one of only two eukaryotic organisms (the other being the protozoan Trypanosoma cruzi) that have been demonstrated to lack ODC activity. As ODC is a key enzyme in polyamine biosynthesis, Arabidopsis is reliant on the additional arginine decarboxylase (ADC; EC 4.1.1.9) pathway, found only in plants and some bacteria, to synthesize putrescine. By using site-directed mutants of the Arabidopsis ADC1 and heterologous expression in yeast, we show that ADC, like ODC, is a head-to-tail homodimer with two active sites acting in trans across the interface of the dimer. Amino acids K136 and C524 of Arabidopsis ADC1 are essential for activity and participate in separate active sites. Maximal activity of Arabidopsis ADC1 in yeast requires the presence of general protease genes, and it is likely that dimer formation precedes proteolytic processing of the ADC pre-protein monomer.     

Pyruvoyl-dependent arginine decarboxylase from Methanococcus jannaschii: crystal structures of the self-cleaved and S53A proenzyme forms

The three-dimensional structure of pyruvoyl-dependent arginine decarboxylase from Methanococcus jannaschii was determined at 1.4 A resolution. The pyruvoyl group of arginine decarboxylase is generated by an autocatalytic internal serinolysis reaction at Ser53 in the proenzyme resulting in two polypeptide chains. The structure of the nonprocessing S53A mutant was also determined. The active site of the processed enzyme unexpectedly contained the reaction product agmatine. The crystal structure confirms that arginine decarboxylase is a homotrimer. The protomer fold is a four-layer alphabetabetaalpha sandwich with topology similar to pyruvoyl-dependent histidine decarboxylase. Highly conserved residues Asn47, Ser52, Ser53, Ile54, and Glu109 are proposed to play roles in the self-processing reaction. Agmatine binding residues include the C terminus of the beta chain (Ser52) from one protomer and the Asp35 side chain and the Gly44 and Val46 carbonyl oxygen atoms from an adjacent protomer. Glu109 is proposed to play a catalytic role in the decarboxylation reaction.     

Purification of S-adenosylmethionine decarboxylase from soybean.

S-Adenosylmethionine decarboxylase (EC 4.1.1.19) was purified to homogeneity from the cytosol of soybean (Glycine max) axes by ammonium sulfate fractionation, DEAE-Sepharose and methylglyoxalbis(guanylhydrazone)-Sepharose 6B chromatographies. The enzyme was free from diamine oxidase activity. The molecular weight of the enzyme estimated by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis was 66,000. The Km value for S-adenosylmethionine was 0.26 mM. The optimum pH and temperature were 7.5 and 40 degrees C. Neither putrescine nor Mg2+ affected the enzyme activity, but the enzyme was inhibited by spermidine, spermine, methylglyoxalbis(guanylhydrazone), sodium borohydride and phenylhydrazine. Agmatine was a novel inhibitor which inhibited S-adenosylmethionine decarboxylase and arginine decarboxylase, preventing the accumulation of decarboxylated S-adenosylmethionine and putrescine, respectively.     

Arginine decarboxylase as the source of putrescine for tobacco alkaloids

The putrescine which forms a part of nicotine and other pyrrolidine alkaloids is generally assumed to arise through the action of ornithine decarboxylase (ODC). However, we have previously noted that changes in the activity of arginine decarboxylase (ADC), an alternate source ofputrescine, parallel changes in tissue alkaloids, while changes in ODC activity do not. This led us to undertake experiments to permit discrimination between ADC and ODC as enzymatic sources of putrescine destined for alkaloids. Two kinds of evidence presented here support a major role for ADC in the generation ofputrescine going into alkaloids: (a) A specific ‘suicide inhibitor’ of ADC effectively inhibits the biosynthesis of nicotine and nornicotine in tobacco callus, while the analogous inhibitor of ODC is less effective, and (b) the flow of ¹⁴C from uniformly labelled arginine into nicotine is much more efficient than that from ornithine.     

Arginine decarboxylase is a component activity of the multifunctional enzyme putrescine synthase in cucumber seedlings

A homogenous PreParation of Putrescine synthase, the versatile multifunctional enzyme involved in agmatine →Putrescine conversion inCucumis sativus was found to catalyze enzymatic decarboxylation of arginine also. Similarly, the Purified arginine decarboxylase mediated the comPonent as well as the comPlete set of couPled reactions harboured by Putrescine synthase. Both the enzyme PreParations exhibited identical electroPhoretic and chromatograPhic behaviour and were immunologically indistinguishable. All the enzymic activities are stabilized concurrently by feeding arginine to the intact seedlings. Therefore, it is concluded that the multifunctional Putrescine synthase inCucumis sativus seedlings also harbours arginine decarboxylase activity unlike its counterPart inLathyrus sativus.     

Expression of human arginine decarboxylase, the biosynthetic enzyme for agmatine

Agmatine, an amine formed by decarboxylation of L-arginine by arginine decarboxylase (ADC), has been recently discovered in mammalian brain and other tissues. While the cloning and sequencing of ADC from plant and bacteria have been reported extensively, the structure of mammalian enzyme is not known. Using homology screening approach, we have identified a human cDNA clone that exhibits ADC activity when expressed in COS-7 cells. The cDNA and deduced amino acid sequence of this human ADC clone is distinct from ADC of other forms. Human ADC is a 460-amino acid protein that shows about 48% identity to mammalian ornithine decarboxylase (ODC) but has no ODC activity. While naive COS-7 cells do not make agmatine, these cells are able to produce agmatine, as measured by HPLC, when transfected with ADC cDNA. Northern blot analysis using the cDNA probe indicated the expression of ADC message in selective human brain regions and other human tissues.     

The therapeutic and nutraceutical potential of agmatine,and its enhanced production using Aspergillus oryzae

Agmatine, a natural polyamine produced from arginine by arginine decarboxylase, was first discovered in 1910, but its physiological significance was disregarded for a century. The recent rediscovery of agmatine as an endogenous ligand for α2-adrenergic and imidazoline receptors in the mammalian brain suggests that this amine may be a promising therapeutic agent for treating a broad spectrum of central nervous system-associated diseases. In the past two decades, numerous preclinical and several clinical studies have demonstrated its pleiotropic modulatory functions on various molecular targets related to neurotransmission, nitric oxide synthesis, glucose metabolism, polyamine metabolism, and carnitine biosynthesis, indicating potential for therapeutic applications and use as a nutraceutical to improve quality of life. An enzymatic activity of arginine decarboxylase which produces agmatine from arginine was low in mammals, suggesting that a large portion of the agmatine is supplemented from diets and gut microbiota. In the present review, we focus on and concisely summarize the beneficial effects of agmatine for treating depression, anxiety, neuropathic pain, cognitive decline and learning impairment, dependence on drugs, and metabolic diseases (diabetes and obesity), since these fields have been intensively investigated. We also briefly discuss agmatine content in foodstuffs, and a simple approach for enhancing agmatine production using the filamentous fungus Aspergillus oryzae, widely used for the production of various Asian fermented foods.     

Stereochemistry of reactions catalysed by arginine decarboxylase and ornithine decarboxylase

The decarboxylations of l-arginine, catalysed by arginine decarboxylase (EC 4.1.1.19) and of l-ornithine, catalysed by ornithine decarboxylase