Amino Acid composition of germinating cotton seeds

Total and free amino acid composition of germinating cotton seeds (Gossypium hirsutum L.) was determined. The germinating seeds were separated into cotyledon and developing axis fractions daily and the composition of each tissue was summed to get the whole seed composition. By separating the developing seeds into these two tissue fractions, and determining total and free amino acids, a balance sheet was developed for each amino acid. This technique allowed changes in distribution with time of each amino acid to be followed in each tissue. Data for total content and amount in protein of each amino acid are presented. Asparagine increased in the whole seed, and most of this increase was found in the free pool of the developing axis. Other amino acids (e.g. arginine, glutamic acid) increased in the free pool but showed an over-all decrease, indicating that they were being metabolized. Amino acid contents of storage and nonstorage protein isolates were determined.     

Amino acid and protein changes in the haemolymph of developing fourth instar Chironomus tentans

The concentration of free amino acids, total soluble protein, and haemoglobin in the haemolymph of fourth instar Chironomus tentans was investigated.The concentration of the free amino acid pool increases between the early (15.7 mM/l) and mid-(33.9 mM/l) fourth larval stages followed by a decline during the late (16.9 mM/l) fourth larval period. Alanine, serine, and the amides of aspartic acid and glutamic acid are the predominant free amino acids at all stages. Physiological fluid analysis of late fourth instar haemolymph detected 32 ninhydrin positive components including 18 common amino acids plus homoarginine, ornithine, citrulline, β-alanine, α-aminoadipic acid, α-aminoisobutyric acid, and sarcosine.The concentration of total soluble protein steadily increases during fourth instar larval development to a maximum of $\text{9.3}\text{g}\text{100}\text{ml}$ followed by a decline during the pharate pupal period. A similar pattern of variation occurs in haemoglobin content which comprises from 51 to 66% of Chironomus tentans haemolymph protein.The mM percentage of individual amino acids of total haemolymph protein varies little during the fourth instar. At all stages alanine and aspartic acid are the predominant amino acids.     

Effect of sound and Lenzites-decayed wood on the amino acid composition of Reticulitermes flavipes

In hydrolysates of the eastern subterranean termite, Reticulitermes flavipes, the most abundant protein amino acids (μmoles) were glycine, alanine, and glutamic acid; the least abundant were methionine and histidine. Sawdust from both sound and Lenzites trabea-decayed sapwood blocks of sugar maple, loblolly pine, and slash pine was force-fed to termites. A diet of decayed rather than sound wood had little effect on protein amino acid composition of the termites; glycine content varied the most. In contrast, diet affected the free amino acid composition. Except for glutamic acid, the major protein amino acids of the termites were not the predominant free amino acids. Tyrosine and histidine were relatively more abundant as free than as protein amino acids. Greatest differences in protein amino acid compositions of sound and decayed wood were in contents of glycine, leucine, lysine, and arginine.     

Amino Acid Metabolism of Lemna minor L. : I. Responses to Methionine Sulfoximine

When Lemna minor L. is supplied with the potent inhibitor of glutamine synthetase, methionine sulfoximine, rapid changes in free amino acid levels occur. Glutamine, glutamate, asparagine, aspartate, alanine, and serine levels decline concomitantly with ammonia accumulation. However, not all free amino acid pools deplete in response to this inhibitor. Several free amino acids including proline, valine, leucine, isoleucine, threonine, lysine, phenylalanine, tyrosine, histidine, and methionine exhibit severalfold accumulations within 24 hours of methionine sulfoximine treatment. To investigate whether these latter amino acid accumulations result from de novo synthesis via a methionine sulfoximine insensitive pathway of ammonia assimilation (e.g. glutamate dehydrogenase) or from protein turnover, fronds of Lemna minor were prelabeled with [¹⁵N]H₄⁺ prior to supplying the inhibitor. Analyses of the ¹⁵N abundance of free amino acids suggest that protein turnover is the major source of these methionine sulfoximine induced amino acid accumulations. Thus, the pools of valine, leucine, isoleucine, proline, and threonine accumulated in response to the inhibitor in the presence of [¹⁵N]H₄⁺, are ¹⁴N enriched and are not apparently derived from ¹⁵N-labeled precursors. To account for the selective accumulation of amino acids, such as valine, leucine, isoleucine, proline, and threonine, it is necessary to envisage that these free amino acids are relatively poorly catabolized in vivo. The amino acids which deplete in response to methionine sulfoximine (i.e. glutamate, glutamine, alanine, aspartate, asparagine, and serine) are all presumably rapidly catabolized to ammonia, either in the photorespiratory pathway or by alternative routes.     

Amino acid budgets in three aphid species using the same host plant

Abstract. Nutrient provisioning in aphids depends both on the composition of ingested phloem sap and on the biosynthetic capabilities of the aphid and its intracellular symbionts. Amino acid budgets for three aphid species, Rhopalosiphum padi (L.), Schizaphis graminum (Rondani) and Diuraphis noxia (Mordvilko), were compared on a single host plant species, wheat Triticum aestivum L. Ingestion of amino acids from phloem, elimination of amino acids in honeydew, and the content of amino acids in aphids tissue were measured. From these values, ingestion rates were estimated and compared to honeydew and to estimated composition of aphid proteins. Ingestion rate was lowest in D. noxia due to low growth rate and low honeydew production; intermediate in S. graminum due to higher growth rate and intermediate honeydew production; and highest in R. padi , which had the highest rates for both variables. Both D. noxia and S. graminum induced increases in the amino acid content of ingested phloem. These changes in phloem content, combined with differences in ingestion rates, resulted in large differences among aphids in estimated rates of ingestion of individual amino acids. In honeydew, most essential amino acids were found in low amounts compared with the amounts ingested, especially for methionine and lysine. A few amino acids (arginine, cystine, histidine and tryptophan) were more abundant in honeydew of some aphids, suggesting oversupply. Aphid species differed in the composition of free amino acids in tissue but showed very similar composition in protein, implying similar requirements among the aphids. In R. padi and D. noxia , most essential amino acids were ingested in amounts insufficient for growth, implying dependence on symbiont provisioning. In S. graminum , most amino acids were ingested in amounts apparently sufficient for growth.     

Alteration in the Amino Acid Content of Yeast During Growth Under Various Nutritional Conditions

Yeast cells grown under optimal and suboptimal concentrations of biotin were analyzed for the amino acid content of their soluble pool and cellular protein. Optimally grown yeast cells exhibited a maximum amino acid content after 18 hr of growth. Biotin-deficient cells were depleted of all amino acids at 26 and 43 hr, with alanine, arginine, aspartate, cysteine, glutamate, isoleucine, leucine, lysine, methionine, serine, threonine, and valine being present in less than half the concentration observed in biotin-optimal cells. At early time intervals, the amino acid pool of biotin-deficient yeast contained lower concentrations of all amino acids except alanine. After more prolonged incubation, several amino acids accumulated in the pool of biotin-deficient yeast, but citrulline and ornithine accumulated to appreciable levels. The addition of aspartate to the growth medium resulted in a decrease in the amino acid content of biotin-optimal cells but caused a marked increase in the concentration of amino acids in biotin-deficient cells. The pools of biotin-deficient yeast grown in the presence of aspartate displayed a marked reduction in every amino acid with the exception of aspartate itself. These data provide evidence that the amino acid content of yeast cells and their free amino acid pools are markedly affected by biotin deficiency as well as by supplementation with aspartate, indicating that aspartate plays a major role in the nitrogen economy of yeast under both normal as well as abnormal nutritional conditions.     

6种石斛属植物氨基酸组成及营养价值评价

为比较不同种石斛的氨基酸组成差异,作出营养评价,使用氨基酸自动分析仪检测6种石斛的氨基酸含量,分析其必需氨基酸组成成分,并通过计算氨基酸评分(AAS)、化学评分(CS)及氨基酸比值系数分(SRCAA)等非生物指标进行营养评价。结果表明:石斛中氨基酸种类齐全、营养丰富,均含有被检测的17种氨基酸。6种石斛氨基酸总量存在差异,介于3.58%～8.09%。必需氨基酸组成总含量介于42.62%～47.23%之间,明显高于WHO/FAO模式值(35.00%),其中,化学评分显示其限制氨基酸均为半胱氨酸和蛋氨酸。6种石斛药用氨基酸含量相差较大(2.24%～5.37%),其中蜻蜓石斛含量最高,为含量最低细叶石斛的2.40倍,但在总氨基酸中所占的比例基本一致(60.96%～66.47%)。对6种石斛进行聚类分析可分为4类,其中杓唇石斛、叠鞘石斛、细叶石斛可视为高品质蛋白质种。综上所述,石斛氨基酸具有重要的营养价值,6种石斛氨基酸差异显著。该结果可为石斛营养价值评价、品种选育及药用开发提供理论指导。     

Amino Acid Metabolism of Lemna minor L. : III. Responses to Aminooxyacetate

Aminooxyacetate, a known inhibitor of transaminase reactions and glycine decarboxylase, promotes rapid depletion of the free pools of serine and aspartate in nitrate grown Lemna minor L. This compound markedly inhibits the methionine sulfoximine-induced accumulation of free ammonium ions and greatly restricts the methionine sulfoximine-induced depletion of amino acids such as glutamate, alanine, and asparagine. These results suggest that glutamate, alanine, and asparagine are normally catabolized to ammonia by transaminase-dependent pathways rather than via dehydrogenase or amidohydrolase reactions. Aminooxyacetate does not inhibit the methionine sulfoximine-induced irreversible deactivation of glutamine synthetase in vivo, indicating that these effects cannot be simply ascribed to inhibition of methionine sulfoximine uptake by amino-oxyacetate. This transaminase inhibitor promotes extensive accumulation of several amino acids including valine, leucine, isoleucine, alanine, glycine, threonine, proline, phenylalanine, lysine, and tyrosine. Since the aminooxyacetate induced accumulations of valine, leucine, and isoleucine are not inhibited by the branched-chain amino acid biosynthesis inhibitor, chlorsulfuron, these amino acid accumulations most probably involve protein turnover. Depletions of soluble protein bound amino acids are shown to be approximately stoichiometric with the free amino acid pool accumulations induced by aminooxyacetate. Aminooxyacetate is demonstrated to inhibit the chlorsulfuron-induced accumulation of α-amino-n-butyrate in L. minor, supporting the notion that this amino acid is derived from transamination of 2-oxobutyrate.     

Changes in accumulation of seed nitrogen compounds in maize under conditions of sulphur deficiency

Maize ( Zea mays L., hybrid INRA 260) was grown in the greenhouse with mineral nutrition of different sulphate concentrations. Mature seeds from these plants were compared for their free amino acid and protein N forms. For the most S-deficient sample, the Asx (asparagine + aspartic acid) content increased by 30% as compared with control, while methionine and cysteine decreased (by 25 and 30%, respectively), as well as glycine, lysine, histidine, arginine and tryptophan. In seeds lowest in S the non-protein N to total N ratio was 77% higher than in the control. Free asparagine dominated in starved seeds (50 mol % of total free amino acids) and was ten-fold more concentrated than in the control, where proline was the predominant free amino acid. Thus the Asx of non-protein N reached 28% of the total mol Asx of the whole starved seed. Altered S nutrition had virtually no effect on the amino acid composition of the main protein fractions, but it significantly changed their ratios. Zeins, which are poor in S-containing amino acids, showed 25% higher level than in seeds supplied with normal S. As a counterbalance, two glutelin subfractions rich in S-containing amino acids, decreased by 36–71% under limiting S nutrition.
It is concluded that the plant reacts against S deficiency by modifying its N metabolism. Significant accumulation occurred of free asparagine, which is the main form of N transportation. The biosynthesis of seed storage protein occurred through the accumulation of the highest possible protein quantity allowed by the available S-containing amino acids, i.e. proteins low in S-containing amino acids were preferentially synthesized.     

Alleviation of Changes in Protein Metabolism in NaCl-Stressed Wheat Seedlings by Thiamine

NaCl-stress induced a pronounced suppression in growth of wheat seedlings. The most abundant amino acids (cysteine, arginine, methionine) constituting about 55 % of total free amino acid content in control wheat were reduced in 100 mM NaCl-treated plants. However, valine, isoleucine, aspartic acid and proline accumulated in response to NaCl stress and NaCl-treated wheat seedlings showed 1.6 fold increase in total free amino acids compared to the control. Addition of 2 [micro ]M thiamine alleviated the effects of NaCl on the amino acid composition and the amount of total free amino acids decreased to that in the control. Content of 26 kDa protein increased in NaCl-treated plants, stimulation was more pronounced in roots than in shoots. In contrast, the contents of 13 and 20 kDa proteins decreased. After addition of thiamine, the 24 kDa protein, which disappeared with NaCl treatment, has been initiated again. Moreover, thiamine treatment stimulated the accumulation of the 20 kDa protein.     

Free Amino Acid and Storage Protein Composition of Soybean Fruit Explants and Isolated Cotyledons Cultured with and without Methionine

Thein vitroculture of immature soybean cotyledons (in direct contact with the medium) and immature fruit explants (stem dipping into the medium) on a defined medium containing glutamine and sulphate as sole sources of N and S for 7 d led to rates of growth and reserve protein accumulation close to, or greater than, those occurringin situ. Supplementation of the medium with 8.4 mMmethionine had little effect on growth and protein accumulation of the cotyledons in the explant system, but did result in significant increases in the isolated cotyledon system. Methionine suppressed the synthesis of the 7S β-subunit in both systems. The free amino pool of the cotyledons increased more than three-fold when methionine was present in the explant medium. In the isolated cotyledon system, the basal medium alone caused a large increase (over 30-fold) in the free amino acid fraction, but methionine resulted in an even greater increase (over 50-fold). In both systems the expansion involved a very large increase in the methionine pool, but many other amino acids also showed large increases. Specific effects of methionine on individual amino acids were more clear in the explant system, where its presence resulted in marked increases in serine, alanine and asparagine. The data show that an abnormal situation arises on feeding with methionine, a fact to be considered before attributing effects on growth and protein synthesis directly to methionine.     

Metabolic activation and hepatotoxicity. Effect of cysteine, N-acetylcysteine, and methionine on glutathione biosynthesis and bromobenzene toxicity in isolated rat hepatocytes.

Freshly isolated rat hepatocytes contained a high level (30–40 nmol/10⁶ cells) of reduced glutathione (GSH) which decreased steadily upon incubation in an amino acid containing medium lacking cysteine and methionine. This decrease in GSH level was prevented, and turned into a slight increase, when either cysteine, N-acetylcysteine, or methionine was also present in the medium. The amino acid uptake into hepatocytes was more rapid with cysteine than with methionine. Cystine was not taken up, or taken up very slowly, by the cells and could not be used to prevent the decrease in GSH level which occurred in the absence of cysteine and methionine. The level of GSH in hepatocytes freshly isolated from rats pretreated with diethylmaleate was markedly decreased (to ~5 nmol/10⁶ cells) but increased rapidly upon incubation of the cells in a medium containing amino acids including either cysteine, N-acetylcysteine, or methionine. Again, cysteine was taken up into the cells more rapidly than methionine. The rate of uptake of cysteine was moderately enhanced in hepatocytes with a lowered level of intracellular GSH as compared to cells with normal GSH concentration. Exclusion of glutamate and/or glycine from the medium did not markedly affect the rate of resynthesis of GSH by hepatocytes incubated in the presence of exogenously added cysteine or methionine. Incubation of hepatocytes with bromobenzene in an amino acid-containing medium lacking cysteine and methionine resulted in accelerated cell damage. Addition of either cysteine, N-acetylcysteine, or methionine to the medium caused a decrease in bromobenzene toxicity. The protective effect was dependent, however, on the time of addition of the amino acid to the incubate; e.g., the effect on bromobenzene toxicity was greatly reduced when either cysteine or methionine was added after 1 h of preincubation of the hepatocytes with bromobenzene as compared to addition at zero time. This decrease in protective effect in bromobenzene-exposed cells was related to a similar decrease in the rate of uptake of cysteine and methionine into hepatocytes preincubated with bromobenzene. The rate of uptake, and incorporation into cellular protein, of leucine was also markedly inhibited in hepatocytes preincubated with bromobenzene. In contrast, there was no measurable change in the rate of release of leucine from cellular protein as a result of incubation of hepatocytes with bromobenzene. It is concluded that the presence of cysteine, N-acetylcysteine, or methionine in the medium protects hepatocytes from bromobenzene toxicity by providing intracellular cysteine for GSH biosynthesis and suggested that an inhibitory effect on amino acid uptake may contribute to the cytotoxicity of bromobenzene in hepatocytes.     

Amino acid requirements of the mosquito Culex pipiens: asparagine essential

When any of the ten “rat essential” amino acids was omitted singly from a fully-defined synthetic dietary medium, newly-hatched Culex pipiens larvae were unable to develop to the second instar. With proline omitted, development was greatly retarded and survival to the adult stage reduced. Without aspargine (but not aspartic acid) growth and development ceased in most individuals before larval-pupal ecdysis, and no adults were obtained. These twelve amino acids are considered nutritionally essential for this mosquito. With glycine omitted singly, development was markedly retarded, but survival to the adult stage was not affected; thus this amino acid is required for good growth, but these experiments do not demonstrate it as essential. Single omission of alanine, aspartic acid, cysteine, glutamic acid or amide, serine or tyrosine had virtually no effect on development and they are therefore considered nutritionally non-essential. With diets containing the twelve culex-essential amino acids only, very little development occurred, but augmentation with either glycine or serine allowed growth and development almost as good as with the complete amino acid mixture. Augmentation of the essential twelve with alanine, cysteine, glutamic acid/amide, or tyrosine singly failed to improve development. The requirement for dietary asparagine shown by these studies appears to be unique among insects so far studied. In particular, another mosquito, Aedes aegypti, has no such requirement.     

Amino Acids from Heterodera glycines

Amino acids emitted and extracted from surface-sterilized larvae and adults of Heterodera glycines were identified by paper chromatography and quantitatively analyzed by column chromatography. Five amino acids (alanine, aspartic acid, glutamic acid, glycine and serine) were emitted by H. glycines larvae and eight others (asparagine, glutamine, leucine/isoleucine, lysine, methionine sulfoxide, threonine, tyrosine, valine/methionine) were found in extracts from crushed larvae.In addition to the amino acids emitted or extracted from larvae, four others were emitted by adults (γ-aminobutyric acid, histidine, phenylalanine, and proline). Four different amino acids (arginine, cystathionine, hydroxyproline, and ornithine) were found only in the extract from crushed adults. Greater quantities of alanine, aspartic acid and glycine were emitted than could be detected in nematode extracts suggesting selective emission.Subsamples of nematode populations were taken from growing plants 19, 26, 33, and 40 days after inoculation and extracted to determine whether changes in specific amino acid content correlated with aging. Proline content shifted most, increasing from 4.1% to 21.5% of the total amino acid complement from the 19th to the 40th days.     

Protein content and amino acids composition of bee-pollens from major floral sources in Al-Ahsa,eastern Saudi Arabia

Protein content and amino acids composition of bee-pollens from major pollen floral sources in Al-Ahsa, Saudi Arabia were determined to investigate the nutritive value of pollen protein relative to requirements of honeybees and adult humans. The major pollen sources were alfalfa, date palm, rape, summer squash, and sunflower. Bee-pollens from alfalfa and date palm showed high content of crude protein and amino acid concentrations. Bee-pollen from sunflower had low content of those components. Eighteen amino acids were found in bee-pollens from the five major floral sources. The highest concentrations of individual amino acids valine, leucine, isoleucine, phenylalanine and proline were obtained from alfalfa bee-pollen; lysine, arginine, cysteine, tryptophan and tyrosine from date palm; methionine, histidine, glycine and alanine from summer squash; threonine, serine and glutamic acid from sunflower; and aspartic acid from rape bee-pollen. The amino acid composition obtained from sunflower bee-pollen showed the lowest concentrations of the essential amino acids: isoleucine, leucine, methionine, phenylalanine and valine. Apart from methionine, arginine and isoleucine, the essential amino acids of bee-pollen from alfalfa, date palm, summer squash and rape exceeded the honeybees’ requirements. Methionine was the limiting amino acid in bee-pollens from the five selected sources. Concentrations of essential amino acids in the tested bee-pollens were variable and significantly correlated to their botanical origin of pollen. Bee-pollens from alfalfa, date palm and summer squash was found to be rich source of protein and amino acids for bees and for humans.     

Mutational Bias Plays an Important Role in Shaping Longevity-Related Amino Acid Content in Mammalian mtDNA-Encoded Proteins

During the course of evolution, amino acid shifts might have resulted in mitochondrial proteomes better endowed to resist oxidative stress. However, owing to the problem of distinguishing between functional constraints/adaptations in protein sequences and mutation-driven biases in the composition of these sequences, the adaptive value of such amino acid shifts remains under discussion. We have analyzed the coding sequences of mtDNA from 173 mammalian species, dissecting the effect of nucleotide composition on amino acid usages. We found remarkable cysteine avoidance in mtDNA-encoded proteins. However, no effect of longevity on cysteine content could be detected. On the other hand, nucleotide compositional shifts fully accounted for threonine usages. In spite of a strong effect of mutational bias on methionine abundances, our results suggest a role of selection in determining the composition of methionine. Whether this selective effect is linked or not to protection against oxidative stress is still a subject of debate.     

Amino acid composition of vascular sap of maize ear peduncle

The amino acid composition of the vascular sap of a high lysine maize mutant was determined during kernel development. With the exception of proline and cystine, all amino acids that occur in the endosperm were found in the vascular sap of the ear peduncle. Glutamine is the major amino acid transported to the endosperm varying from 30.6 to 20.6 μmol at 7 and 42 days after pollination, respectively. Aspartic acid, the second most important nitrogen form translocated to the seeds, was ca 10 μmol% during kernel filling. Glutamine and arginine content decreased with maturity, while valine, methionine, isoleucine, leucine, tyrosine and phenylalanine increased with kernel development. The remaining N forms were constant during endosperm growth.     

Anisakis physeteris: Amino acids in body tissues

The whole cuticle, perienteric fluid, and reproductive organs of Anisakis physeteris Baylis, 1923 from the sperm whale were analyzed for amino acid composition. Amino acid nitrogen per total nitrogen accounted for 96.2% in the cuticle. Corresponding values of TCA supernatant and precipitated fractions of perienteric fluid and reproductive organ were obtained. Proline, glycine, and arginine occurred abundantly in cuticle hydrolyzate. Lysine, glutamic acid, glycine, valine, leucine and histidine occounted for approximately 62% of total nitrogen in reproductive organ fractions. Differences were observed in the amounts of certain amino acids present in corresponding tissue of males and females as well as different tissue fractions from nematodes of the same sex, e.g., more serine in male cuticle than female, more proline in the protein of female reproductive organs than male; methionine present in cuticular protein and reproductive organ protein and nonprotein fractions but absent in perienteric fluid, and cystine in cuticular protein but not the other fractions. Untreated Anisakis hemolymph showed at least a 2-fold increase in ammonia in analyses made 2 and 10 days after bleeding. The relatively large ammonia content could have been in part the result of functional ornithine cycle and amino acid oxidase activity in vivo.     

Transgenic approaches to improve the nutritional quality of plant proteins

Summary Plant proteins, when used as dietary protein, are generally incomplete in nutrition due to their deficiency in several essential amino acids, for example, lysine and tryptophan in cereals and methionine and cysteine in legumes. Attempts to breed crops with increased levels of lysine and methionine have been less than satisfactory. Modern biotechnology offers alternative approaches for rectifying this nutrition deficiency. In the past decade, several transgenic strategies aimed at modifying the amino acid composition of plant proteins and enhancing the content of specific essential amino acid(s) for nutrition improvement have been developed and tested. These include synthetic proteins, modification of protein sequences, over-expression of heterologous or homologous proteins, and metabolic engineering of the free essential amino acid pool and protein sink. The progress and potential of these approaches and studies are reviewed. As plant proteins are the primary source of all dietary protein consumed by humans and animals and are inexpensive to produce in comparison with meat, improving their quality will make a significant contribution to our future food needs. The research and development in this area of interest is making promising progress towards this endeavor.     

Amino acid supply and protein metabolism in Ehrlich ascites tumour cells. 1. Identification of limiting amino acids

The uptake and intracellular accumulation of an amino acid mixture by incubated Ehrlich ascites tumour cells was studied. The composition of the amino acid mixture simulated that of mouse intraperitoneal fluid and amino acid uptake was studied over a range of concentrations between 0.0 (no added amino acids) and 10.0-times intraperitoneal concentrations. For most amino acids uptake into cells and intracellular accumulation occurred as concentrations were increased up to 6.0-times the intraperitoneal concentrations; further increases in external amino acid concentrations did not increase concomitantly with intracellular concentrations. These data, when analysed indicated a net protein synthetic rate of 20% d-1 and that the rate of protein synthesis may be limited by the availability of the amino acids lysine, threonine and methionine.