Free Amino Acid Composition of Phloem Sap and Growing Fruit of Lycopersicon esculentum

Pure phloem sap of tomato leaves was collected by stylectomy.Glutamine and glutamate were the predominant free amino acidstranslocated by the phloem stream. In developing fruits glutaminecontent increased significantly, reaching 35% of the total freeamino acids. Comparison in the amino acid composition betweenthe two tissues are discussed. (Received October 6, 1997; Accepted January 27, 1998)     

Correlation between Protein Body Formation and Storage Protein Accumulation in Soybean Cotyledons

At 20 days after flowering (DAF), the 7S α' and α subunits began to accumulate. At 25 DAF, the 7Sβ, l1SA and llSB subunits appeared. Five days later, the 11SA-4 subunit was present During the period of 25–55DAF, the storage protein content continued to increase. From 55 to 63 DAF, there was a decrease in the synthetic rate of the storage proteins. Comparing these results with the two paths of protein body formation reported previously, we draw the conclusion that the protein bodies developed from vacuoles contained not only the 7S bm also the lis proteins in soybean cotyledon cells.     

Genomic Regions Associated with Amino Acid Composition in Soybean

Soybean [Glycine max (L.) Merr.] is the single largest source of protein in animal feed. However, few studies have been conducted to evaluate genomic regions controlling amino acid composition in soybean. It is important to study the genetics of amino acid composition to achieve improvements through breeding. The objectives of this study were to determine the ratios between essential to non-essential (E:NE) and essential to total (E:T) amino acids, and to identify genomic regions controlling essential and non-essential amino acid composition in soybean seed. To achieve these objectives, 101 F₆-derived recombinant inbred lines (RIL) developed from a cross of N87-984-16 × TN93-99 were used. Ground soybean seed samples were analyzed for amino acids using a near infrared spectroscopy (NIRS) instrument. A significant (p < 0.01) difference among the RIL was found for amino acid composition. Heritability estimates on an entry mean basis ranged from 0.13 for His to 0.67 for Tyr. A total of 94 polymorphic simple sequence repeat (SSR) molecular genetic markers were screened in DNA from progenies. Single factor ANOVA was used to identify candidate quantitative trait loci (QTL), which were then confirmed by QTL Cartographer. At least one QTL for each amino acid was detected in this population. QTL linked to molecular markers Satt143, Satt168, Satt203, Satt274 and Satt495 were associated with most of the amino acids. Phenotypic variation explained by an individual QTL ranged from 9.4 to 45.3%. QTL detected for amino acids in soybean in this experiment are expected to be useful for future breeding programs targeting development of improved soybean amino acid composition for human and animal nutrition.     

Developmental Changes in the Free Amino Acid Pool and Total Protein Amino Acids of Pea Cotyledons (Pisum sativum L.)

Changes in the levels of twenty-two free amino acids and in the amino acid composition of the total protein were measured throughout the development of cotyledons of a dwarf garden pea, Pisum sativum cv Greenfeast, grown in a constant environment. A sensitive double-isotope dansylation technique was used. Fresh weight, dry weight, and protein content were also followed. Twenty of the amino acids showed synchronous changes in levels, giving a developmental pattern containing four peaks; major peaks occurred very early and very late in development. The amino acid composition of the total protein, which was always very different from that of the free amino acid pool, showed early changes to one consistent with the final storage protein composition of the seed. These changes included a 50% drop in methionine content and a 70% rise in cysteine. While the maximum free methionine level occurred early in development, that of cysteine was late.     

高纬度地区大豆蛋白含量及氨基酸组分的表型鉴定与聚类分析

对国内外高纬度地区的240份大豆种质资源的蛋白及16个氨基酸组分进行测定,通过遗传多样性、因子和聚类分析,进行了表型鉴定及基因型分类。结果表明,供试大豆种质蛋白及氨基酸组分变异较丰富,遗传多样性程度较高。根据因子分析,将筛选到的3个公因子进行聚类分析,可将供试种质资源分为7类。蛋白含量从高到低的顺序为类群Ⅶ>类群Ⅵ>类群Ⅴ>类群Ⅱ>类群Ⅰ>类群Ⅲ>类群Ⅳ。12个氨基酸组分的变化趋势与蛋白一致。类群Ⅶ和类群Ⅵ为高蛋白遗传群体,可作为高蛋白基因聚合育种的亲本材料。通过前期分析,筛选到24份高蛋氨酸资源,包括有公野04L-141、龙品03-311、Proto、和龙油太、猫眼豆、茶色豆、紫花2号、东农48等,可为高蛋氨酸种质创新提供材料基础。     

Changes in Amino Acid Composition during Germination of Soybean

As a fundamental approach to the problem of amino acid metabolism in soybean, changes in content of twenty-three free amino acids, two acidic peptides, ammonia, ethanolamine, urea and seventeen total amino acids of cotyledon, hypocotyl and root of soybean during germination were determined with an amino acid analyzer. Glycine Max M. var. T201 (non-nodule-forming) and Glycine Max M. var. T202 (nodule-forming) were used for this experiment. The content and composition of free and total amino acids of cotyledon in both T201 and T202 differ from those of other tissues in any stage of germination. However, no significant difference between these two varieties of soybean has been recognized in patterns of free and total amino acids changes during germination.

In dry bean and initial stage of germination a relatively large unknown peak appeared and disappeared thereafter when the change in free amino acid content during germination of soybean was analyzed with amino acid analyzer. From various tests on the unknown peak, it became obvious that the peak was consisted of two peptides, γ-glutamyl-tyrosine and γ-glutamylphenylalanine, which were discovered in soybean by Thompson et al. in 1962. The content of these peptides did not change during the first 20 hours of germination, but they decreased rapidly thereafter and disappeared after 70 hours.     

Changes in Amino Acid Composition during Germination of Soybean

α- and γ-Glutamylaspartic acids were detected in acidic fraction of soybean seedling. R_F values of both peptides were consistent with those of authentic samples with several solvent systems.     

Changes in Amino Acid Composition during Germination of Soybean

Malonogalactan, a malonylated polysaccharide (—74° (c=1.6, H₂O)) produced by Penicillium citrinum, consisted of d-galactose and malonic acid in the approximate molar ratio of 3:1. Molecular weight of the demalonylated galactan (-99° (c=4.6, H₂O)) was about 40,000. From the data regarding optical rotation, nuclear magnetic resonance spectrum, infrared spectrum, glycosidase susceptibility, periodate oxidation, Smith degradation, methylation and acid hydrolysis, the possible structure of the Penicillium malonogalactan is deduced as follows: A galactan, 1,5-β-galactofuranoside polymer esterified with malonic acid at the position of 2 or 3.     

氨基酸组成对蛋白质耐热性的影响

为了研究一级结构对蛋白质耐热性的影响,利用软件DNAMAN对16个家族32种蛋白质序列进行了氨基酸含量分析,并统计分析了氨基酸组成对蛋白质耐热性的影响。通过比较同一家族的高低温蛋白质序列及16个家族中所有高温和低温蛋白质序列中氨基酸含量的变化可以推断(从低温到高温)：Ser、Cys．含量降低显著,Arg、Ile、Pro含量升高显著。由此可知高温蛋白质倾向于含有疏水性氨基酸而避免亲水性氨基酸。     

Amino Acid Composition of Ferredoxin and Rubredoxin Isolated from Desulfovibrio gigas

The composition of ferredoxin and rubredoxin from Desulfovibrio gigas was determined. Ferrodoxin contained five cysteines, two methionines, and four irons. The rubredoxin was not unique.     

Storage Protein Composition of Soybean Cotyledons Grown In Vitro in Media of Various Sulfate Concentrations in the Presence and Absence of Exogenous l-Methionine

Supplemental methionine in a complete culture medium increased the methionine content of the protein fraction of cultured soybean (Glycine max L. Merrill) cotyledons (Thompson, Madison, Muenster 1981 Phytochemistry 20: 941-945). To explain the observed increase in protein methionine, we have measured the amounts and subunit compositions of 7S and 11S storage proteins and determined the amino acid compositions of the three major protein fractions (2-5S, 7S, 11S) of seeds developed on plants and of cultured cotyledons grown in the presence or absence of supplemental l-methionine. Development of cultured cotyledons was representative of development of seeds on plants. The ratios of 11S to 7S proteins, the subunit contents, and amino acid compositions of their storage protein fractions were similar, but not identical. Supplemental methionine increased the mole percent methionine in each of the three protein fractions of cultured cotyledons and changed the amounts of several other amino acids. Supplemental methionine inhibited expression of the 7S β-subunit gene. Concomitant with the absence of the β-subunit, which contains no methionine, was an increase in the ratio of 11S to 7S proteins, and an increase in the methionine content of the subunits composing these fractions. Inhibition of β-subunit gene expression by methionine in cultured cotyledons provides a reproducible, easily controlled system for the study of eucaryotic gene expression.     

根据蛋白质的氨基酸组成实现其快速鉴定

常规进行蛋白质鉴定的方法是测定其氨基酸顺序,它需要蛋白质顺序分析仪,对蛋白质的纯度要求高,费时和花费大,与之相比,蛋白质的氨基酸组成和分子量是容易实验测定的。本文描述了一个基于蛋白质的组成和分子量进行其快速鉴定的方法。其基本出发点是,通过统计蛋白质序列数据库中每个序列的氨基酸组成和分子量,得到一个含蛋白质长度、组成和分子量的数据库,将靶蛋白质的组成等数据与该数据库进行对比,可以检出组成和分子量与之接近的蛋白质。从而对该蛋白质进行初步鉴定。在有些情况下,甚至能相当准确地确定靶蛋白质与数据库中的某个（些）蛋白质相关。根据这一原理本文设计了根据氨基酸组成检索蛋白质组成数据库的程序,通过对胰岛素原、细胞肿瘤抗原Ｐ５３和泛肽等多种蛋白质的组成分析,证实根据氨基酸组成能较好地进行蛋白质鉴定。     

Protein Content and Amino Acid Composition of Certain Fungi Evaluated for Microbial Protein Production

The protein and total amino acid contents of four mycelial fungal strains and one yeast were approximately the same for cultures harvested in the mid-log and early stationary growth phases. It was found that Fusarium oxysporum and Fusarium moniliforme contained approximately 30% more protein and total amino acids than Aspergillus niger. The amino acid composition of mycelial protein compares favorably with that of British Petroleum yeast protein Toprina produced commercially on hydrocarbon substrates. Fusarium spp. may be suitable for commercial production of microbial protein, especially when low-cost agricultural or industrial waste products are readily available as energy sources. Genetic manipulation of these fungi, such as induction of mutant strains through irradiation, may be desirable to obtain a mycelial product of improved yield and/or quality.     

SuccSite: Incorporating Amino Acid Composition and Informative k-spaced Amino Acid Pairs to Identify Protein Succinylation Sites

Protein succinylation is a biochemical reaction in which a succinyl group (-CO-CH2-CH2-CO-) is attached to the lysine residue of a protein molecule. Lysine succinylation plays important regulatory roles in living cells. However, studies in this field are limited by the difficulty in experimentally identifying the substrate site specificity of lysine succinylation. To facilitate this process, several tools have been proposed for the computational identification of succinylated lysine sites. In this study, we developed an approach to investigate the substrate specificity of lysine succinylated sites based on amino acid composition. Using experimentally verified lysine succinylated sites collected from public resources, the significant differences in position-specific amino acid composition between succinylated and non-succinylated sites were represented using the Two Sample Logo program. These findings enabled the adoption of an effective machine learning method, support vector machine, to train a predictive model with not only the amino acid composition, but also the composition of k-spaced amino acid pairs. After the selection of the best model using a ten-fold cross-validation approach, the selected model significantly outperformed existing tools based on an independent dataset manually extracted from published research articles. Finally, the selected model was used to develop a web-based tool, SuccSite, to aid the study of protein succinylation. Two proteins were used as case studies on the website to demonstrate the effective prediction of succinylation sites. We will regularly update SuccSite by integrating more experimental datasets. SuccSite is freely accessible at http://csb.cse.yzu.edu.tw/SuccSite/.     

Amino Acid Composition of Bulk Protein of Euglena Grown in Waste Water

The amino acid content of bulk protein in a sewage-grown Euglena sp. was examined. Concentrations of the essential amino acids, threonine, histidine, tryptophan, and valine, were similar to those found in other algae. The concentration of alanine was much higher. Methionine was not found at all, proline only in traces, and other amino acids at low concentrations. These results indicate that the amino acid content of bulk protein of the species of Euglena studied resembles that of plants far more closely than that of animals.     

Control of Aromatic Amino Acid Biosynthesis in Cultured Plant Tissues: Effect of Intermediates and Aromatic Amino Acids on Free Levels

Shikimate, anthranilate, indole, l -tryptophan, phenylpyruvate, l -p henylalanine, p-hydroxyphenylpyruvate or l -tyrosine were added to suspension-cultured Nicotiana tabacum (tabacco) and Daucus carota (carrot) tissues and incubated for 24 hours. Uptake of each compound was substantial as measured by its decrease in the medium. The levels of free tryptophan, phenylalanine and tyrosine were determined in the tissues after the 24 hours incubation. Shikimate did not change the aromatic animo acid levels in carrot tissue, but did increase all three in tobacco (3-fold or more), indicating a less stringent feedback control in tobacco. Anthranilate and indole increased the tissue tryptophan levels in both species by at least 17-fold, showing that the flow from anthranilate and indole to tryptophan was apparently unhindered by enzymatic control mechanisms. When tryptophan levels were elevated in both carrot and tobaccotissues by anthranilate, indole or tryptophan addition, there was also an increase in free phyenylalanine and tyrosine. This might be due to the reversal of phenylalanine and tyrosine feedback inhibition of chorismate mutase by the high tryptophan in the tissue. Chorismate mutase activity in tobacco crude extracts could be inhibited by 66–90% by 1 mM phenylalanine and /or tyrosine. Tryptophan at 1 mM stimulated the enzyme activity by 1/3 and completely reversed the phenylalanine and/or tyrosine inhibition of enzyme activity. Chorsimate mutase activity amino acids under a variety of conditions. Phenylpyruvate increased the phenylalanine levels and p-hydroxyphenylpyruvate increased the tyrosine levels in carrot and tobacco tissues indicating that there was no feedback control of the last step in phenylalanine and tyrosine biosynthesis.