聚酰胺薄膜层析及其用于蛋白质化学分析

聚酰胺薄膜层析是1966年后发展起来的一种新层析技术。特别在用于分析氨基酸衍生物——DNS-氨基酸、DNP-氨基酸和PTH-氨基酸时,具有灵敏度高、分辨力强、速度快、操作方便等优点,超过了过去分析这类化合物使用的纸层析、纸电泳、硅胶薄板层析等方法。在蛋白质化学结构分析中,聚酰胺薄膜层析结     

柱前和柱后衍生高效液相色谱分析氨基酸方法进展与评述

柱前和柱后衍生高效液相色谱分析氨基酸方法进展与评述刘惠文中国林业科学研究院分析中心北京１００９１自从１９５８年Ｓｐａｃｋｍａｎｓｔｅｉｎ和Ｍｏｏｒｅ建立了最初的氨基酸分析技术以来,随着高效液相色谱的出现和发展,氨基酸分析方面的探讨也更加广泛和深入［１...     

不同产地天麻氨基酸的含量测定

氨基酸用途广泛,人们对氨基酸质量和品种的要求也越来越高.我国氨基酸从无到有,到现在的大市场份额,经历了五十多年的发展历程,其中包括谷氨酸、赖氨酸、苏氨酸和蛋氨酸在内的大品种氨基酸已经发展成为全球的主要供应品种,部分小品种氨基酸已形成自己的优势.绿色制造和现代生物技术的应用将成为氨基酸行业发展的主流.本文重点论述了我国氨基酸品种的现状、水平以及未来发展方向.     

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。     

异硫氢酸苯酯法(PITC)分析氨基酸

<正> 自从1958年Moore等人建立氨基酸测定方法以来,氨基酸分析方法发展迅速,日新月异。除采用阳离子交换柱,茚三酮柱后反应测定氨基酸方法外,70年代起,各种采用柱前衍生,反相色谱法分离测定氨基酸方法相继问世,为氨基酸分析开辟了新的领域。异硫氢酸苯酯(PITC)法作为其中一种,以其灵敏度高(可检测,1Pmal),分析速度快(水解液为10分钟,生理体液40～60分     

D-氨基酸检测方法研究进展

自然界中存在20种基本氨基酸,除甘氨酸外都有两种互成镜像的对映体D-型和L-型。长期以来,人们认为D-氨基酸仅存在于微生物中,参与构成细胞壁肽聚糖层。但随着分析方法的发展,人们相继在多种生物体中(包括人体)发现了多种D-氨基酸,D-氨基酸才引起人们的高度重视,并意识到它们在医药、农药和食品等的组成中起着重要作用。D-氨基酸检测方法还需要不断发展与完善,以便更好地分析和认识D-氨基酸的重要作用。主要介绍酶法、气相层析法、高效液相法及生物传感器等检测方法。     

利用串联质谱鉴定氨基酸突变的生物信息学算法

氨基酸突变能够改变蛋白的结构和功能,影响生物体的生命过程.基于串联质谱的鸟枪法蛋白质组学是目前大规模研究蛋白质组学的主要方法,但是现有的质谱数据鉴定流程为了提高鉴定结果的灵敏度往往会有意压缩数据库中的氨基酸突变信息.因此,如何挖掘数据中的氨基酸突变信息成为当前质谱数据鉴定的一个重要部分.当前应用于氨基酸突变鉴定的串联质谱鉴定方法大致可以分为3大类:基于序列数据库搜索的方法、基于序列标签搜索的算法以及基于图谱库搜索的算法.本文首先详细介绍了这3种氨基酸突变鉴定算法,并分析了各种方法的特点和不足,然后介绍了氨基酸突变鉴定的研究现状和发展方向.随着基于串联质谱的蛋白质组学的不断发展,蛋白序列中的氨基酸突变信息将被更好地解析出来,从而得以深入探讨由氨基酸突变引起的蛋白结构和功能改变,为揭示氨基酸突变的生物学意义奠定基础.     

氨基酸生产的代谢工程研究进展与发展趋势

氨基酸发酵是我国发酵工业的支柱产业,近年来,随着代谢工程的快速发展,氨基酸的代谢工程育种蓬勃发展。传统的正向代谢工程、基于组学分析与计算机模拟的反向代谢工程以及借鉴自然进化的进化代谢工程,都有越来越多的应用。在氨基酸的工业生产中涌现出了一系列具有高效生产、抗逆性强等优良性状的菌株。日益剧烈的市场竞争对菌株的选育提出了新的要求,如开发高附加值氨基酸品种、菌株代谢的动态调控、适应新工艺的要求等。文中介绍了氨基酸生产相关的代谢工程研究进展以及未来的发展趋势。     

微生物传感器及其在氨基酸分析中的应用

介绍了微生物传感器的发展状况、制作方法,综述了微生物传感器在氨基酸分析中的应用,引文献４２篇。     

淀粉酶的同源性研究

对α－淀粉酶、麦芽四糖淀粉酶和葡萄糖淀粉酶进行的氨基酸序列比较表明,它们之间的氨基酸等同性相当低。本文在对这三种淀粉酶的氨基酸残基进行疏水性分析的基础上,采用国际上近期发展的疏水簇分析方法对这三种淀粉酶的氨基酸序列进行了二维描述,其结果清楚地显示了它们之间的同源性差异。这也被测定的三维结构所证实。这一研究结果表明,与蛋白质三维结构密切相关的蛋白质的二级结构及其相互配置主要取决于蛋白质序列中不同类型氨基酸残基的排列。     

ABRF ESRG 2005 study: identification of seven modified amino acids by Edman sequencing.

Identification of modified amino acids can be a challenging part for Edman degradation sequence analysis, largely because they are not included among the commonly used phenylthiohydantion amino acid standards. Yet many can have unique retention times and can be assigned by an experienced researcher or through the use of a guide showing their typical chromatography characteristics. The Edman Sequencing Research Group (ESRG) 2005 study is a continuation of the 2004 study, in which the participating laboratories were provided a synthetic peptide and asked to identify the modified amino acids present in the sequence. The study sample provided an opportunity to sequence a peptide containing a variety of modified amino acids and note their retention times relative to the common amino acids. It also allowed the ESRG to compile the chromatographic properties and intensities from multiple instruments and tabulate an average elution position for these modified amino acids on commonly used instruments. Participating laboratories were given 2000 pmoles of a synthetic peptide, 18 amino acids long, containing the following modified amino acids: dimethyl- and trimethyl-lysine, 3-methyl-histidine, N-carbamyl-lysine, cystine, N-methyl-alanine, and isoaspartic acid. The modified amino acids were interspersed with standard amino acids to help in the assessment of initial and repetitive yields. In addition to filling in an assignment sheet, which included retention times and peak areas, participants were asked to provide specific details about the parameters used for the sequencing run. References for some of the modified amino acid elution characteristics were provided and the participants had the option of viewing a list of the modified amino acids present in the peptide at the ESRG Web site. The ABRF ESRG 2005 sample is the seventeenth in a series of studies designed to aid laboratories in evaluating their abilities to obtain and interpret amino acid sequence data.     

乙醇系统HPLC分析PTC－氨基酸的条件优化

首次报道用乙醇系统分析ＰＴＣ－氨基酸的新方法中各衍生物获得最佳分离的建立过程。ＰＴＣ－氨基酸衍生后溶于Ａ溶液,然后进样于４μｍＮｏｖａＰａｋＣ１８柱（３．９ｍｍ×１５０ｍｍ）。系统的优化步序包括全面调控流动相的ｐＨ值与ＴＥＡ浓度、乙醇梯度程序、柱温等诸多影响ＨＰＬＣ色谱行为的因素。最适条件为：Ａ溶液含０．１４ＭＧ酸钠、０．７５ｍｌ／ＬＴＥＡ、ＰＨ６．３５;Ｂ溶液为１００％乙醇;柱温３０℃。通过优化的乙醇梯度最终在约４４ｍｉｎ内将１５种ＰＴＣ－氨基酸很好地分离。此法可用于替代代表新科技水平的ＰＴＣ－氨基酸乙腈分析系统。     

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。     

黄粉虫复合氨基酸的提取及氨基酸虫酒的制作

本研究在对黄粉虫幼虫所含的复合氨基酸3种提取方法及其所制成的氨基酸虫酒进行分析比较的基础上,得出最佳方案和数据：用酶解法处理虫浆可得到占干虫重31.5％酶解滤粉和31.06％复合氨基酸粉,是用水解法提取出复合氨基酸粉的2.1倍。在虫体原料重量相同的条件下,酶解滤粉酒中复合氨基酸浓度分别是水解滤粉酒和虫粉酒中复合氨基酸浓度的2.7和13倍。若制成氨基酸浓度为5mp／mL的虫酒,仅需幼虫21.5g,折合成用虫成本费0.5元左右。     

EDMAN SEQUENCING RESEARCH GROUP 2004 STUDY: MODIFIED AMINO ACIDS IN EDMAN SEQUENCING

Edman degradation sequencing relies on comparing high-performance liquid chromatography retention times of the sample phenylthiohydantoin amino acids with phenylthiohydantoin amino acid standards. The elution characteristics of the twenty common amino acids have been well characterized, which aids in making confident assignments. Modified amino acids may present more of a challenge since they are not part of the commonly used standards and because the protein sequencer analyst may not have experience with them. Laboratories requesting a sample were sent a tube containing approximately 775 pmoles of a 20-amino-acid synthetic peptide composed of several modified amino acids that may be found in proteins or are generated during sample preparation. In addition to filling in an assignment sheet, which included retention times and peak areas, participants were asked to provide specific details about the parameters used for the sequencing run. References for some of the modified amino acid elution characteristics were provided and the participants had the option of viewing a list of the modified amino acids present in the peptide at the Edman Sequencing Research Group website (ESRG). The goal of the study consisted of two parts: assessment of the ability to correctly assign all the amino acids in the peptide, including the modified amino acids; and the collection and compiling of elution time characteristics of modified amino acids for instruments used in the study. The resulting compilation of the modified amino acid elution times and running conditions will be accessible at the Association of Biomolecular Resource Facilities (ABRF) ESRG website for future reference. The ABRF ESRG 2004 sample is the 16th in a series of studies designed to aid laboratories in evaluating their abilities to obtain and interpret amino acid sequence data.     

福建产太子参氨基酸成分分析

采用日立L8800全自动高速氨基酸分析仪,从福建柘荣产太子参中检出18种氨基酸,全氨基酸总质量分数为77.7g.kg-1,其中精氨酸(Arg)高达20.8 g.kg-1;此外,还发现太子参中含有丰富的γ-氨基丁酸,质量分数高达16.5 g.kg-1。采用RT-HPLC(柱前衍生-反相液相色谱分离)检测质量分数为20.5 mg.kg-1,验证了HPCEC(离子交换色谱分离-柱后衍生法)氨基酸自动分析结果。     

血浆中氨基酸检测技术探讨

在血浆中检测游离氨基酸含量是临床诊断的重要手段之一。本文讨论了液相色谱法、氨基酸分析法、液质联用技术等几种血浆中氨基酸的检测技术,并介绍这些技术在氨基酸检测中的应用,为建立血浆中游离氨基酸的分析方法提供参考依据。     

Functional analysis of three amino acid residues of purR re-pressor, Trp147, Gln-218 and Gln-292 in Salmonella typhi-murium

The amber mutation sites of 6 purR(am) mutants were determined bycloning and DNA sequencing. The results showed that the mutations were distributed at three different sites in PurR coding region, G721(→A), C933(→T) and C1155(→T), which respectively turn Trp-147,Gln-218 and Gln-292 of PurR into TAG terminal codon. To determine the effect of the three amino acid residues on regulatory function of PurR protein 5 different kinds of tRNA suppressor genes, Su3, Su4, Su6, Su7 and Su9 were used for creating the PurR protein variants with single amino acid substitution. The results indicated that Cys, Glu, Gly, His and Arg which substituted Trp-147 respectively all could not recover the regulation function of PurR. It confirmed that Trp-147 is a critical amino acid for the PurR function. Gln-292 substituted respectively by the same amino acids also could not recover the PurR function, demonstrating that Gln-292 is also an important amino acid residue in PurR.     

非天然氨基酸3-(4-噻唑基)-D,L-丙氨酸盐酸盐的合成

合成了非天然氨基酸 3- ( 4 -噻唑基 ) - D,L -丙氨酸盐酸盐以及三个中间体 ,其结构分别通过红外光谱、核磁共振、元素分析、熔点等测试手段得到确证     

青海循化线辣椒中氨基酸含量分析

采用柱前衍生高效液相色谱法对循化线辣椒中精氨酸、丙氨酸、丝氨酸等17种氨基酸的含量进行测定。结果表明:线辣椒所含17种氨基酸中,必需氨基酸含量合理,利于人体吸收。