冬虫夏草菌菌丝体中的氨基酸及其衍生物的代谢组学分析

为探究冬虫夏草菌（Ophiocordyceps sinensis）菌丝体氨基酸及其衍生物的差异性,采用超高效液相色谱串联质谱（Ultra performance liquid Chromatography, tandem mass spectrometry, UPLC-MS/MS）技术对3株冬虫夏草菌（玉树菌株XSH、达日菌株DR、贵德菌株LJ）菌丝体的氨基酸及其衍生物进行靶向定量检测,利用主成分分析（Principal component analysis, PCA）和正交偏最小二乘判别分析（Orthogonal signal correction and partial least squares-discriminant analysis, OPLS-DA）考察样品分类情况、筛选差异代谢物。结果表明,3株菌株菌丝体中均检测到72种氨基酸及其衍生物,发现除组氨酸外可合成蛋白质的19种氨基酸,菌株XSH的总氨基酸及必需氨基酸绝对含量均高于其他菌株。菌株XSH相比菌株DR和菌株LJ分别含有29种和28种差异氨基酸及其衍生物,菌株DR相比菌株LJ含有10种差异氨基酸及其衍生物,此外,3株菌株菌丝体含有3个共有的差异氨基酸及其衍生物,分别为高精氨酸、N-乙酰-L-谷氨酰胺、肌氨酸。京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes,KEGG）通路富集分析显示,差异氨基酸及其衍生物在精氨酸和脯氨酸代谢通路中更活跃,不同产区冬虫夏草菌株的氨基酸含量可能受精氨酸和脯氨酸代谢通路的影响。基于研究结果发现不同产区的冬虫夏草菌株的氨基酸及其衍生物存在较大的差异。     

含硫氨基酸衍生物的研究进展

含硫氨基酸衍生物主要由三个常见的含硫氨基酸———蛋氨酸 ,半胱氨酸 ,胱氨酸衍生转化而来 ,可以在体内参与转甲基 ,转硫基等生化过程 ,同时也具有抗氧化 ,清除自由基等功能 ,在医药和食品中具有重要的应用价值。含硫氨基酸衍生物产品众多 ,其中最具代表性的是N 乙酰 L 半胱氨酸 ,谷胱甘肽 ,腺苷蛋氨酸。本文就这三个衍生物的研究进展作一简要综述。     

氨基酸酰胺类化合物的合成

阐述了氨基酸氨基保护的常用方法和试剂,氨基酸酰胺类化合物合成的基本原理和方法以及在合成中需要注意的问题。重点阐述了氨基酸酰胺类化合物的合成机理和合成方法。展望了氨基酸酰胺类衍生物的合成方向。     

pH值对高效液相(OPA法)测定氨基酸的影响

<正> 前言氨基酸的分离是在强阳离子中变换柱子通过pH梯度进行的。洗脱出来的各种氨基酸再与OPA反应生成衍生物,最后应用萤光检测分离出的各种氨基酸。其反应如下:     

氨基酸技术发展及新产品开发

<正>氨基酸及其衍生物具有非常重要的生理功能。氨基酸工业是发酵工业的支柱产业之一,其产品有着广泛的应用和巨大的市场。近些年,氨基酸工业的发展日新月异,各种氨基酸生产的新菌种、新工艺和新技术层出不穷,这为氨基酸工业的进一步发展提供了巨大的动力。主要介绍氨基酸代谢工程的技术发展和氨基酸深层次加工及新产品开发进展。     

相思子属三种药材中的氨基酸分析比较

利用全自动氨基酸分析仪和高效液相色谱仪分析比较相思子属三种药材中氨基酸成分及含量。结果9个样品都检测出17种氨基酸和2种色氨酸衍生物。结论:不同样品中的氨基酸含量有所差异,但其中门冬氨酸、谷氨酸、亮氨酸和赖氨酸的含量相对较高。     

氨基酸的气相色谱(续三)

<正> 4.6.为GC选择酰基氨基酸酯(参看第7.2及7.5节)。对于GC分析来说,特殊衍生物的选定决定于一些相关的因素:诸如制备简单,高产率,稳定性以及衍生物的挥发性等。还要选择适于它们分辨力的层析柱和层析条件。有几个小组比较了几类酰基化氨基酸及其酯的挥发性。测定了一些N—TFA—氨基酸的融点,更适当的升华温度以及蒸汽压;在0.02—0.06Torr(毫米汞柱)真空下,几种氨基酸在70—80℃之间     

高灵敏显色降解试剂4-N，N-二甲氨基偶氮苯-4′-异硫氰酸酯(DABITC)在蛋白质顺序分析中的应用

一、序言 Edman降解是蛋白质顺序分析最有效的方法。它由三步反应组成,首先,异硫氰酸苯酯(PITC)在碱性pH下与肽或蛋白质的N-端氨基酸反应生成苯基氨荒酰(PTC)衍生物,然后在酸性pH下生成比原先的肽少一个氨基酸的肽与苯基噻唑啉酮(PTZ)衍生物,后者在酸性     

亮氨酸脱氢酶研究进展及其工业应用

微生物脱氢酶催化羰基不对称还原制备光学纯氨基酸及其衍生物具有非常大的优势。亮氨酸脱氢酶能选择性地催化α-酮酸,氨化还原得到α-氨基酸及其衍生物。本文综述了亮氨酸脱氢酶的来源,理化性质,底物特异性,酶基因工程菌构建等方面的内容及研究进展。从辅酶再生策略,酶膜反应器两方面讨论了其工业化应用,并展望了今后的发展前景。     

保护性氧化盐酸水解法分析蛋白质水解氨基酸

本文介绍了在保护剂存在下,通过过甲酸氧化,再以盐酸水解,在50分钟的分析循环时间内准确地分析17种氨基酸及三种含硫氨基酸衍生物的分析方法。     

High yielding synthesis of N-ethyl dehydroamino acids

Recently we reported the use of a sequence of alkylation and dehydration methodologies to obtain N-ethyl-α, β-dehydroamino acid derivatives. The application of this N-alkylation procedure to several methyl esters of β,β-dibromo and β-bromo, β-substituted dehydroamino acids protected with standard amine protecting groups was subsequently reported. The corresponding N-ethyl, β-bromo dehydroamino acid derivatives were obtained in fair to high yields and some were used as substrates in Suzuki cross-coupling reactions to give N-ethyl, β,β-disubstituted dehydroalanine derivatives. Herein, we further explore N-ethylation of β-halo dehydroamino acid derivatives using triethyloxonium tetrafluoroborate as alkylating agent, but substituting N,N-diisopropylethylamine for potassium tert-butoxide as auxiliary base. In these conditions, for all β-halo dehydroamino acid derivatives, reactions were complete and the N-ethylated derivative could be isolated in high yield. This method was also applied for N-ethylation of non-halogenated dehydroamino acids. Again, with all compounds the reactions were complete and the N-ethyl dehydroamino acid derivatives could be isolated in high yields. Some of these N-ethyl dehydroamino acid methyl ester derivatives were converted in high yields to their corresponding acids and coupled to an amino acid methyl ester to give N-ethyl dehydrodipeptide derivatives in good yields. Thus, this method constitutes a general procedure for high yielding synthesis of N-ethylated dehydroamino acids, which can be further applied in peptide synthesis.     

Antimutagenicity of mono-, di-, and tricaffeoylquinic acid derivatives isolated from sweetpotato (Ipomoea batatas L.) leaf

The caffeoylquinic acid derivatives, 3-mono-O-caffeoylquinic acid (chlorogenic acid, ChA), 3,4-di-O-caffeoylquinic acid (3,4-diCQA), 3,5-di-O-caffeoylquinic acid (3,5-diCQA), 4,5-di-O-caffeoylquinic acid (4,5-diCQA) and 3,4,5-tri-O-caffeoylquinic acid (3,4,5-triCQA), and caffeic acid (CA) were isolated from the sweetpotato (Ipomoea batatas L.) leaf. We examined the antimutagenicity of these caffeoylquinic acid compounds to promote new uses of the sweetpotato leaf. These caffeoylquinic acid derivatives effectively inhibited the reverse mutation induced by Trp-P-1 on Salmonella typhimurium TA 98. The antimutagenicity of these derivatives was 3,4,5-triCQA > 3,4-diCQA = 3,5-diCQA = 4,5-diCQA > ChA in this order. There was no difference in the antimutagenicity of all dicaffeoylquinic acid derivatives. A comparison of the activities and structures of these compounds suggested that the number of caffeoyl groups bound to quinic acid played a role in the antimutagenicity of the caffeoylquinic acid derivatives. The sweetpotato leaves contained distinctive polyphenolic components with a high content of mono-, di-, and tricaffeoylquinic acid derivatives and could be a source of physiological functions.     

N-hydroxybenzenecarboximidic acid derivatives: a new class of nitroxyl-generating prodrugs.

On the basis of the propensity of Piloty's acid to generate nitroxyl (HNO), we previously prepared a number of N,O-bisacylated Piloty's acid derivatives and showed that such prodrugs underwent a disproportionation reaction following ester hydrolysis to give an unstable intermediate that hydrolyzed to nitroxyl. To expand the versatility of this series, we desired some mixed N,O-diacylated Piloty's acid derivatives and devised a synthetic route to them. Such efforts led us, serendipitously, to a new series of heretofore unreported nitroxyl-generating compounds. Thus, benzohydroxamic acid was acylated on the hydroxylamino oxygen and the resulting product converted to its sodium salt. Treatment of this salt with arenesulfonyl chorides would be expected to give the mixed N,O-diacylated derivatives of Piloty's acid. However, the products obtained were the isomeric carboximidic acid derivatives whose structures were deduced from the IR and (13)C NMR spectral frequencies associated with the sp(2) carbons. The structures were verified by analysis of the X-ray crystal structure of a prototype compound of this series. When incubated with porcine liver esterase or mouse plasma, these N-acyloxy-O-arenesulfonylated benzenecarboximidic acid derivatives liberated HNO, measured as N(2)O, as well as the expected arenesulfinic acid and benzoic acid. Alkaline hydrolysis also produced N(2)O, but the major products were the arenesulfonic acid and benzohydroxamic acid. Thus, these N-hydroxybenzenecarboximidic acid derivatives represent a new series of nitroxyl prodrugs that require enzymatic bioactivation before nitroxyl can be liberated.     

Changes in bulk protein of tobacco leaves on aerobic autolysis: Hydrogenation studies and identification of bound quinic acid

During aerobic autolysis and in commercial curing, the bulk proteins of tobacco leaves become coupled with quinic acid, presumably in consequence of coupling of chlorogenic acid congeners with lysine ε-NH₂ groups. Quinic acid derivatives, prepared from acid hydrolysates of such altered proteins, were identified by GC-MS. Such proteins were also hydrogenated over Rh/Al₂O₃ with a view to stabilizing the hypothetical linkages. Difficulties in removing contaminant Al had to be overcome. Evidence was then obtained (by GLC of derivatives) for several components, in acid hydrolysates of hydrogenated altered proteins, which were neither normal hydrogenation products of the common amino acids nor derivatives of quinic acid. Details of the chromatograms and mass spectra of quinic acid derivatives are provided in a supplementary publications.     

Terahertz Time-Domain Spectroscopy of Four Hydroxycinnamic Acid Derivatives

The well-resolved absorption spectra of the hydroxycinnamic acid (HCA) derivatives, caffeic acid, ferulic acid, sinapic acid and chlorogenic acid, were measured over the frequency region from 0.3 to 2.0 THz at 294 K with terahertz time-domain spectroscopy (THz-TDS). Theoretical calculation was applied to assist the analysis and assignment of the individual THz absorption spectra of the HCA derivatives with density functional theory (DFT). The distinctive spectral features were originated from the collective motion of molecules held together by hydrogen bonds. The real and imaginary parts of dielectric function of the four HCA derivatives were also obtained.     

Synthesis of new glycyrrhetinic acid (GA) derivatives and their effects on tyrosinase activity

To synthesize glycyrrhetinic acid (GA) derivatives (3, 4, 5, 10, 13, 14, 15, and 16), we first removed the ketonic group in the C-11 position, and the carboxylic function at the C-30 position was kept intact, reduced to an alcohol, or transformed to an aldehyde corresponding derivatives 10 and 13. Glycyrrhetinic acid (GA) derivatives (3, 4, 5, 15, and 16) were coupled with 4-amino piperpyridine derivatives (12 and 14) and 4-fluorobenzyl bromide at C-30 carboxylic acid position of glycyrrhetinic acid. In subsequent tyrosinase assays, we found that GA derivatives 4, 5, and 16 were not active at early time points, but strongly inhibited tyrosinase activity at late time points. Of the GA derivatives examined, derivative 5 was most active, with an IC₅₀ value of 50 μM after 2 h reaction. IC₅₀ values of derivatives 4 and 16 were 120 and 170 μM, respectively. Further kinetic data indicated that these derivatives are slow-binding inhibitors of tyrosinase. The time-dependent inhibition was reversed when vitamin C or kojic acid was used, that is, both compounds showed active inhibition at early time points. These results suggest that GA derivatives are much more stable than vitamin C or kojic acid, although their intrinsic inhibitory potentials are relatively low. Higher stability and activity suggest that GA derivative 5 might be a useful candidate for skin whitening.     

Synthesis and dual biological effects of hydroxycinnamoyl phenylalanyl/prolyl hydroxamic acid derivatives as tyrosinase inhibitor and antioxidant

We previously reported that caffeoyl-amino acidyl-hydroxamic acid (CA-Xaa-NHOH) acted as both a good antioxidant and tyrosinase inhibitor, in particular when caffeic acid was conjugated with proline or amino acids having aromatic ring like phenylalanine. Here, various hydroxycinnamic acid (HCA) derivatives were further conjugated with phenylalanyl hydroxamic acid and prolyl hydroxamic acid (HCA-Phe-NHOH and HCA-Pro-NHOH) to study the structure and activity relationship as both antioxidants and tyrosinase inhibitors. When their biological activities were evaluated, all HCA-Phe-NHOH and HCA-Pro-NHOH exhibited enhanced antioxidant activity compared to HCA alone. Moreover, derivatives of caffeic acid, ferulic acid, and sinapic acid inhibited lipid peroxidation more efficiently than vitamin E analogue (Trolox). In addition, derivatives of caffeic acid and sinapic acid efficiently inhibited tyrosinase activity and reduced melanin content in melanocytes Mel-Ab cell.     

Studies on Griseolic Acid Derivatives X. Synthesis and Phosphodiesterase Inhibitory Activity of 2-Substituted Derivatives of Griseolic Acid

Abstract

Griseolic acid derivatives which were modified at the 2-and/or 6-positions were first synthesized from griseolic acid by a ring opening—reclosure reaction of the adenine ring. Among these derivatives, the 2-amino-6-deamino-6-hydroxyl (guanine) derivative showed 3.3 and 45 times stronger inhibitory activity against cAMP and cGMP PDE, respectively, than those of griseolic acid. Structure-activity relationships among these derivatives are also discussed.     

Beta-oxidation of very-long-chain fatty acids and their coenzyme A derivatives by human skin fibroblasts

The beta-oxidation of lignoceric acid (C24:0), hexacosanoic acid (C26:0), and their coenzyme A derivatives was investigated in human skin fibroblast homogenates. The cofactor requirements for oxidation of lignoceric acid and hexacosanoic acid were identical but were different from their coenzyme A derivatives. For example, lignoceric acid and hexacosanoic acid oxidation was strictly ATP dependent whereas the oxidation of the corresponding coenzyme A derivatives was ATP independent. Also the rate of oxidation of coenzyme A derivatives of lignoceric acid or hexacosanoic acid was much higher compared to the free fatty acids. In patients with Zellweger's syndrome, X-linked adrenoleukodystrophy and infantile Refsum's disease, the beta-oxidation of lignoceric and hexacosanoic acids was defective whereas the oxidation of their corresponding coenzyme A derivatives was nearly normal. The results presented in this communication suggest strongly that the beta-oxidation of very-long-chain fatty acids occurs exclusively in peroxisomes. However, the coenzyme A derivatives of very-long-chain fatty acids can be oxidized in mitochondria as well as in peroxisomes. The inability of the mitochondrial system to oxidize free fatty acids may be due to its inability to convert them to their corresponding coenzyme A derivatives. Our results suggest that a specific very-long-chain fatty acyl CoA synthetase may be required for the activation of the free fatty acids and that this synthetase may be deficient in patients with Zellweger's syndrome and possibly X-linked adrenoleukodystrophy, as well. The results presented suggest that substrate specificity and the subcellular localization of the synthetase may regulate the beta-oxidation of very-long-chain fatty acids in the cell.     

Design, synthesis, and characterization of BK channel openers based on oximation of abietane diterpene derivatives

Oxime ether derivatives at the benzylic position of unsubstituted, dichloro, trichloro, and monobromo derivatives of the aromatic C-ring of dehydroabietic acid and podocarpic acid were synthesized and evaluated as BK channel openers in an assay system of CHO-K1 cells expressing hBKα channels. Detailed SAR analysis showed that the oximation was particularly effective in the cases of dehydroabietic acid derivatives, and some of these oxime derivatives showed more potent BK channel activities than the standard compound, NS1619. The present studies provide a new structural basis for development of efficient BK channel openers.