The response of adult rats to varying combinations of dietary lysine and threonine

The response of adult rats fed diets containing amino acid mixtures with varying combinations of lysine and threonine was studied by estimating the change in body weights, organ weights, total nitrogen content of organs and the concentration of free plasma lysine and threonine. Analysis of variance indicates a significant effect of threonine, lysine and a significant interaction due to feeding the two amino acids on the body weights, organ weights and total nitrogen content of various organs. Significant effect on the level of the plasma free amino acid was also shown as a result of feeding the particular amino acid. Results show that in deficient animals the rates of building body protein are low but are higher in the lysine deficient animals than in the threonine deficient and protein deficient animals. The varying rates of exchanging protein between the various organs in response to changing the levels of amino acids in the diet would mean that total change in body protein or total nitrogen balance may not be satisfactory way to determine the protein or amino acid requirements for maintenance. It seems to be necessary to focus down on protein turnover rates in specific tissues.     

Endogenous lysine and other amino acid flows at the terminal ileum of the growing pig (20 kg bodyweight): The effect of protein-free,synthetic amino acid,peptide and protein alimentation

Twenty-four crossbred pigs of 15 kg initial bodyweight were fed four semi-synthetic diets for 10 days according to a completely randomised design. The study aimed to determine the effects of state of body nitrogen balance and the presence of dietary peptides and protein in the digestive tract on the excretion of endogenous amino acids from the ileum of the pig. Endogenous lysine excretion was determined for pigs given a protein-free (PF) diet, an enzymically hydrolysed casein- (EHC), a zein- (ZN) or a synthetic amino acid- (SAA) based diet. Digesta from the EHC-fed animals were centrifuged and ultrafiltered after collection and the precipitate plus retentate fraction was used to determine the endogenous flows. Such processing excludes unabsorbed dietary amino acids from the measure of endogenous loss. ZN is naturally deficient in lysine and tryptophan and these two amino acids were omitted from the synthetic amino acid-based diet to allow direct measurement of endogenous lysine flow. Pigs given the ZN and SAA diets received free lysine and tryptophan orally throughout the study except for the final 2 days of the study, when these amino acids were infused intravenously. Endogenous flows for amino acids other than lysine were determined for pigs given the PF and EHC diets. On the final day of the study the pigs were given their daily dietary allowance hourly and killed 10 h after the start of feeding. Digesta were collected from the terminal ileum (20 cm anterior to the ileo-caecal junction) and endogenous flows were determined by reference to the marker chromic oxide. The mean endogenous ileal lysine flows for the ZN- and EHC-fed pigs were not significantly different (overall mean, 419 mg kg^?1 dry matter intake), but were higher (P < 0.05) than those for the PF- and SAA-fed pigs (overall mean, 268 mg kg^?1 dry matter intake) whose mean flows were not significantly different from each other. The mean endogenous ileal flows for amino acids other than lysine were higher (P < 0.05) for the EHC-fed pigs compared to the animals on the PF diet, except for proline, glycine and arginine. The similar endogenous ileal lysine excretion for pigs receiving a SAA-based diet and in positive body nitrogen balance, and PF-fed pigs in negative body nitrogen balance, indicates that negative body nitrogen balance per se does not lead to a lowered endogenous ileal excretion. It would appear, however, that the presence of dietary peptides or protein in the gut increases amino acid excretion at the terminal ileum above that found with PF or SAA alimentation. Consequently, endogenous ileal amino acid flow in the pig may be underestimated when determined by the traditional PF method.     

酶解脱脂蚕蛹蛋白过程中鲜味规律研究

以缫丝蚕蛹为实验材料,分析碱性蛋白酶和风味蛋白酶复配酶解脱脂蚕蛹蛋白规律。结果表明:酶解脱脂蚕蛹蛋白的最适底物质量浓度为8?g/100?m L、酶解温度为50℃、总酶添加量为1?g/100?g(以脱脂蚕蛹粉计)、酶解时间为9 h;酶解过程中促进了糖的释放,可溶性氮主要是以肽氮形式存在,酶解产物的分子质量主要集中在小于1 k D;在酶解产物中主要的游离氨基酸分别为苏氨酸、组氨酸、亮氨酸、酪氨酸、赖氨酸、半胱氨酸,占总游离氨基酸含量的60%左右,对酶解液的滋味产生一定的影响;在酶解产物中肽类氨基酸主要是鲜甜味氨基酸,占总肽类氨基酸的50%左右,其中亲水性氨基酸与碱性氨基酸含量的比值约为1;通过对酶解产物鲜味的相关性分析,肽类氨基酸的含量与酶解产物的鲜味有较好的线性相关性。     

Nutritive value of oat and rye grain protein as influenced by nitrogen and amino acid composition

In experiments with increasing applications of nitrogen to oats and rye grown in pots, grain with widely varying N-content was obtained. In rye increasing N-concentrations were associated with decreases in the protein of lysine and several other amino acids including threonine, tryptophan, methionine and cystine, whereas glutamic acid, proline and phenylalanine increased. In oat protein the amino acid composition was less affected by increasing N-content in grain. Concentrations of most essential amino acids were higher in oat than in rye protein, especially at high N-levels. The ratio lysine + arginine/proline was higher in oat than in rye protein and did not, as in rye, decrease with increasing N-concentration. Lysine as a percentage of dry matter increased up to the highest N-level in both cereals. In rat feeding experiments true digestibility of the protein of oats and rye increased to the same extent with increasing N-concentration in grain, whereas the biological value of the protein decreased considerably in rye but only slightly in oats. Net protein utilisation decreased with increasing N-content in rye but increased in oats.     

菲律宾蛤仔外套膜液含氮物分布特性及贮藏变化

以活品菲律宾蛤仔外套膜液为研究对象,对外套膜液含氮物以及在活品贮藏过程中的变化进行了分析。鲜活原料分为四个处理组,即对照组、15 ℃室温下的湿藏与干藏组、及4 ℃冷却干藏组。结果显示,外套膜液水分含量高达95%以上;干物质中主要由灰分、蛋白及多糖组成,含量分别为69.41%、15.82%及15.11%;外套膜液抽提氮占总氮70%。与原料组相比,三个贮藏组的总氮、抽提氮、游离氨基酸和总糖有明显升高趋势,总体而言,干藏比湿藏高,冷却干藏比室温干藏高。游离氨基酸分析结果显示,不同贮藏组的各个氨基酸虽存在明显差异,具有呈味功能的谷氨酸、甘氨酸、精氨酸均占总FAA的50%以上;7种必需氨基酸包括苏氨酸、缬氨酸、甲硫氨酸、异亮氨酸、亮氨酸、苯丙氨酸及赖氨酸均有检测,其中赖氨酸含量最高。蛋白态氮的氨基酸组成分析表明,各个贮藏组苏氨酸、缬氨酸、甲硫氨酸、异亮氨酸、亮氨酸、苯丙氨酸及赖氨酸等必需氨基酸含量占总氨基酸的比值在25%~37%。总之,不同的贮藏处置对菲律宾蛤仔含氮物分布均有影响,其中冷却干藏的影响最大。菲律宾蛤仔外套膜液富含蛋白与多糖,其富含呈味FAA及氨基酸的特点,为开发具有营养功能的海鲜调味料提供参考。     

Nutritive value of potato crude protein as influenced by manuring and amino acid composition.

Potatoes were grown in pots and fertilised with varying amounts of nitrogen (as nitrate or ammonium sulphate or urea), phosphorus, potassium and cow manure. The largest yield, 451 g DM per pot, was obtained with nitrogen in the form of urea. N-applications and P- and K-deficiency in the soil increased the total-N content, which varied from 1.14–3.07%. Concentrations of nitrate-N (0.001–0.016%) were negligible. Increasing N-concentrations were associated with decreases in the crude protein of most amino acids including lysine (6.26–4.17%), threonine (4.08–2.90%), methionine (1.98–1.46%) and tryptophan (1.74–0.86%). The concentrations of aspartic and glutamic acids (probably mainly present as their amides) increased. Differences in amino acid composition between boiled and unboiled potatoes were negligible. From 70 to 90% of the decrease in concentration of most essential amino acids could be accounted for by changes in N-content of dry matter. Phosphorus and potassium affected the amino acid composition indirectly through their effects on N-content. In rat feeding experiments increasing N-concentrations in potatoes increased the true digestibility of the crude protein from 79 to 91 but decreased the biological value from 82 to 59. The net protein utilisation did also decrease (from 67 to 54). These relationships were linear below 2.2% N in dry matter. Changes were mainly dependent on variations in N-content. Decreases in essential amino acid indices or chemical scores were closely reflected in the results of the feeding experiments. The true amino acid digestibilities were different for individual amino acids and increased with increasing N-concentration in dry matter.     

CHANGES IN NITROGEN FRACTIONS AND FREE AMINO ACIDS DURING RIPENING OF SPANISH DRIED BEEF "CECINA"

The changes in nitrogen fractions (total nonprotein nitrogen -NPN-, amino acid nitrogen, peptide nitrogen and volatile basic nitrogen) and free amino acids during the ripening of "cecina" (a Spanish dried beef product) were tracked. The amount of total NPN, amino acid nitrogen and volatile basic nitrogen increased during processing, while peptide nitrogen decreased in the last stage. An increase was observed in all amino acids except taurine. In the final product, the most abundant free amino acids were leucine-isoleucine mixture, lysine and proline-methionine mixture. Taking into account the concentrations of the most abundant amino acids detected in cecina and the taste thresholds described for L-amino acids in water, it appears that several amino acids i.e. lysine, alanine, valine and glutamic acid, could contribute to the flavor of this meat product.     

Effects of nitrogen,phosphorus and potassium on amino acid composition and on relationships between nitrogen and amino acids in wheat and oat grain

During two seasons, pot experiments in which increasing applications of nitrogen, phosphorus and potassium were given to wheat and oats produced grain with widely varying N contents. The total amino acid composition of the grain was mainly dependent on N content of grain. Phosphorus and potassium affected the amino acid composition of wheat and oat grain only indirectly through their effects on nitrogen concentration. In wheat increasing grain N% was accompanied by a decrease in the amount (as g per 16 g N) of lysine, threonine, methionine, cystine and an increase in glutamic acid, proline, phenylalanine, and serine contents. Lysine, methionine, and cystine in oats decreased with increasing grain N content, but arginine, aspartic acid, glutamic acid and phenylalanine increased. Changes were less pronounced with oats than with wheat, and concentrations of most essential amino acids were higher in oats. When expressed as % dry matter all amino acids increased with increasing N concentration. Linear regression equations were calculated and significant correlations were found in both crops between concentrations of most amino acids and N content in grain. Some correlations between amino acids in the grain are also given. Correlations between amino acids and nitrogen within a variety were similar despite widely differing yields.     

Milk production,nutrient utilization,and endocrine responses to increased postruminal lysine and methionine supply in dairy cows

The effect of increased postruminal supply of lysine and methionine was investigated in a production trial involving 64 dairy cows in early lactation. Within each of two basal rations, based on either corn silage or grass silage, rations were either naturally deficient in lysine or fortified with 24 g of lysine in a rumen-protected form and naturally deficient in methionine or fortified with 12 g of methionine in a rumen-protected form. The data were analyzed separately for the four lysine and the four methionine treatment groups. Milk production, body weight gain, and plasma concentrations of insulin-like growth factor-I, bovine somatotropin, insulin, glucose, nonesterified fatty acids, and urea were monitored over a 12-wk period. Supplementation with protected methionine led to increases in milk fat and protein contents of 2.4 and 1.8 g/kg of milk, respectively. Supplementation with protected lysine or methionine numerically increased protein yield comparable to values reported in the literature, but the treatment effects were not statistically significant. Efficiency of utilization of absorbed amino acids for milk protein synthesis and efficiency of utilization of metabolizable energy for milk production were not significantly altered in response to increased postruminal lysine and methionine flow, but a numerically increased efficiency of utilization of total amino acids was observed. No significant effect of lysine or methionine supplementation was observed on endocrine parameters nor on plasma metabolite concentrations. However, across treatment groups, high milk yield was correlated with low plasma insulin-like growth factor-I concentrations (r = -0.44) and partially with low plasma nonesterified fatty acids concentration and insulin levels (r = -0.26), while body weight gain was negatively correlated (r = -0.33) with elevated plasma bovine somatotropin concentrations.     

肉类贮藏过程中FAA和BAs变化规律及腐败标识物确定

目的:确定肉类腐败过程蛋白降解产物作为肉类腐败变质标识物的可能性。方法:通过模拟肉类物流运输贮藏过程,利用滴定法、茚三酮柱后衍生法和高效液相色谱法,对贮藏期间肉类挥发性盐基氮,游离氨基酸和生物胺含量进行测定。结果:随着放置时间的增加,某些游离氨基酸(赖氨酸、酪氨酸、精氨酸)含量会发生不同程度的降解,而样品挥发性盐基氮和腐胺,尸胺和酪胺含量会呈不同程度的上升。以挥发性盐基氮为腐败程度参考,进一步对以上各指标进行皮尔逊相关性分析得到各指标与挥发性盐基氮的相关系数。结论:在常温物流条件下,腐胺和酪胺可作为肉类腐败标识物;在冷链物流条件下,酪胺和腐胺可作为猪后腿肌肉腐败标识物,酪胺和尸胺为牛颈部肌肉腐败标识物。      

添加赖氨酸和苏氨酸对小麦面筋蛋白降低大鼠同型半胱氨酸效果的影响

目的在小麦面筋蛋白中添加赖氨酸和苏氨酸,探讨两种限制氨基酸在小麦面筋蛋白中降低大鼠血浆高同型半胱氨酸(Hcy)血症的影响。方法 32只健康Wistar大鼠随机分为4组,分别为大豆分离蛋白组(25S)、酪蛋白组(25C)、小麦面筋蛋白组(29G)、赖氨酸+苏氨酸添加组(29GLT)。各组大鼠分别给予不同饲料喂养14 d后处死,采集血液、肝脏等样品,用于测定生化、酶学指标。结果添加赖氨酸和苏氨酸可有效改善大鼠因摄入小麦面筋蛋白导致的体重、血浆中Hcy的降低,可抑制血浆S-腺苷蛋氨酸(SAM)和肝脏甜菜碱高半胱氨酸甲基转移酶(BHMT)活性的升高,但对胱硫醚β-合成酶(CBS)活性无影响。蛋氨酸含量较低的大豆分离蛋白组,血浆中同型半胱氨酸的含量并未降低。结论小麦面筋蛋白对大鼠血浆同型半胱氨酸浓度的影响可能与其氨基酸的组成特点有关。小麦面筋蛋白的高半胱氨酸和低赖氨酸及低苏氨酸的氨基酸组成是小麦面筋蛋白降低同型半胱氨酸水平的可能因素。     

Effect of the duration of feeding of a protein‐free diet on endogenous ileal nitrogen and amino acid loss in the growing pig

The aim of the study was to determine whether endogenous nitrogen and amino acid excretions at the terminal ileum change over time in the growing pig fed a protein‐free diet for 8 days. Seven entire male pigs with an overall mean live weight of 81.6 kg (SEM 3.3 kg) and surgically implanted post‐valve T caecum cannulas were fed a semi‐synthetic casein‐based diet for 8 days. Food was withheld from the pigs for 24 h, after which they were fed a protein‐free diet for a further 8 days at a rate of 10% of metabolic body weight per day. Chromic oxide was included in the protein‐free diet as an indigestible marker. Ileal digesta were collected continuously from 13:00 to 18:00 h on each day of the experimental period. Endogenous ileal nitrogen flows were determined for each pig each day the protein‐free diet was given, and endogenous ileal amino acid flows for the first and eighth days. There were no significant (P > 0.05) effects of the duration of feeding of the protein‐free diet on endogenous ileal total nitrogen or amino acid flows, except for the amino acids glycine and cysteine, the flows of which significantly decreased over the 8 day period (P < 0.01 and P < 0.05 for glycine and cysteine respectively), from (mean ± SEM) 1639 ± 217 to 892 ± 212 µg g⁻¹ dry matter intake (DMI) for glycine and from 173 ± 13 to 127 ± 19 µg g⁻¹ DMI for cysteine. The relative contributions (moles of each amino acid as a proportion of total moles of amino acids) of threonine, glycine and cysteine decreased significantly (P < 0.05) and that of proline increased significantly (P < 0.05) during the 8 days that the protein‐free diet was fed to the pigs. © 2000 Society of Chemical Industry     

Chemometric analysis of proteolysis during ripening of Ragusano cheese

Chemometric modeling of peptide and free amino acid data was used to study proteolysis in Protected Denomination of Origin Ragusano cheese. Twelve cheeses ripened 3 to 7 mo were selected from local farmers and were analyzed in 4 layers: rind, external, middle, and internal. Proteolysis was significantly affected by cheese layer and age. Significant increases in nitrogen soluble in pH 4.6 acetate buffer and 12% trichloroacetic acid were found from rind to core and throughout ripening. Patterns of proteolysis by urea-PAGE showed that rind-to-core and age-related gradients of moisture and salt contents influenced coagulant and plasmin activities, as reflected in varying rates of hydrolysis of the caseins. Analysis of significant intercorrelations among chemical parameters revealed that moisture, more than salt content, had the largest single influence on rates of proteolysis. Lower levels of 70% ethanol-insoluble peptides coupled to higher levels of 70% ethanol-soluble peptides were found by reversed phase-HPLC in the innermost cheese layers and as the cheeses aged. Non-significant increases of individual free amino acids were found with cheese age and layer. Total free amino acids ranged from 14.3 mg/g (6.2% of total protein) at 3 mo to 22.0 mg/g (8.4% of total protein) after 7 mo. Glutamic acid had the largest concentration in all samples at each time and, jointly with lysine and leucine, accounted for 48% of total free amino acids. Principal components analysis and hierarchical cluster analysis of the data from reversed phase-HPLC chromatograms and free amino acids analysis showed that the peptide profiles were more useful in differentiating Ragusano cheese by age and farm origin than the amino acid data. Combining free amino acid and peptide data resulted in the best partial least squares regression model (R(2) = 0.976; Q(2) = 0.952) predicting cheese age, even though the peptide data alone led to a similarly precise prediction (R(2) = 0.961; Q(2) = 0.923). The most important predictors of age were soluble and insoluble peptides with medium hydrophobicity. The combined peptide data set also resulted in a 100% correct classification by partial least squares discriminant analysis of cheeses according to age and farm origin. Hydrophobic peptides were again discriminatory for distinguishing among sample classes in both cases.     

Varietal differences in the amino acid composition of barley grain during development and under varying nitrogen supply

The amino acid composition of developing grains of the barley varieties Hiproly and Maris Mink was studied from heading to maturity. Hiproly accumulated approximately 50% more nitrogen per seed than Maris Mink, but maintained a balance of amino acids typical of a low nitrogen barley. The effect of varying nitrogen supply on the amino acid composition of Hiproly and a number of commercial varieties was investigated by growing plants in gravel culture in the glasshouse. In the commercial varieties the proportions of glutamic acid, proline and phenylalanine in the protein were positively correlated with protein content but the levels of the essential amino acids, particularly lysine, showed a strong negative correlation with protein content. Hiproly, however, deviated significantly from this negative correlation of the essential amino acids with protein content.     

Sequence in which essential amino acids become limiting for growth of rats fed rations containing cottonseed meal

Two studies were made to determine the sequence in which essential amino acids become limiting for growth of rats fed rations containing 10% protein from cottonseed meal. In the first study, a mixture of six essential amino acids in which cottonseed proteins are deficient, as compared with human milk proteins, was added to the meal. Each of the amino acids was omitted sequentially and the effect on protein efficiency ratio (PER) and growth was studied. In the second study, the amino acids were added to the meal singly and in combinations. Data regarding PER and net protein utilisation (NPU) obtained in the two studies show that lysine is the first limiting amino acid in cottonseed proteins, followed by threonine, isoleucine and sulphur amino acids.     

Protein and free amino acid patterns in maize ensiled with or without urea

Free and protein amino acid contents of whole-plant maize ensiled with or without urea were determined at intervals during the fermentation process. The free amino acid content of both untreated and urea-treated silage increased with time in storage, but the increases were generally larger in the treated silage. The true protein of untreated silage decreased with time in storage. In urea-treated silage many individual amino acids increased considerably. Among those were the essential amino acids isoleucine, lysine, threonine and valine. The result was a net gain of several amino acids in the treated silage while there was a general decline of most amino acids in the untreated silage. Urea-treated silage at day 20 contained 38% more true protein and 150% more free amino acids than did untreated silage. The implications of these changes for dairy cattle nutrition are discussed.     

A study on the ripening of white pickled Brinza cheese made from ewe's milk

The chemical composition, the microbiological feature and the microstructure of young and mature Brinza cheese were investigated. During ripening fat, total nitrogen, soluble nitrogen, non protein nitrogen; peptide nitrogen, amino acid nitrogen and acidity increased while the microbial feature decreased. The amount of free amino acids and free fatty acids increased with storage and the patterns of their distribution changed with the highest levels for lysine, glutamic, oleic, palmatic and myristic acids. The microstructure of fresh Brinza cheese consists of casein micelles joined together to form an open network with clustering of micelles. During ripening, these micelles continued to aggregate and coalesce with increasing fusion process to give more homogeneous and loose structure similar to that in white pickled Domiati cheese.     

Nutritional upgrading of a new soya-protein isolate—SP55

A series of biochemical and nutritional studies was performed in order to establish the reasons for the poor nutritional quality of the new soya-protein isolate SP55. Moist heat treatment which destroyed the substantial residual anti-tryptic activity present in the SP55 fraction, did not improve the nutritive value of the protein as indicated by rat feeding tests. Furthermore, partial in-vitro-enzymic digestion by papain or by pancreatin of SP55 prior to rat feeding did not alter the poor performance of the preparation. Amino acid analysis indicated that the protein was deficient in several essential amino acids. Subsequent rat feeding studies showed that the nutritional quality of the protein can be highly upgraded by supplementation with sulphur-containing amino acids, tryptophan and threonine.     

AMINO ACID COMPOSITION AND NUTRITIONAL IMPLICATIONS OF MILK FERMENTED BY VARIOUS LACTIC CULTURES

A total of nine milk products were analyzed for total and free amino acids and total and nonprotein nitrogen. The products were: (1) whole milk (WM), (2) skim milk (SM), (3) WM fermented by Streptococcus thermophilus, (4) SM fermented by S. thermophilus, (5) WM fermented by Lactobacillus bulgaricus, (6) SM fermented by L. bulgaricus, (7) WM fermented by L. acidophilus, (8) SM fermented by L. acidophilus and (9) commercially prepared buttermilk. Essential amino acid content of the fermented milk was lower than that of the whole or skim milk. Fermentation of milk by lactic cultures decreased the total lysine (except butter milk) and methionine contents of all the samples. Since these two amino acids are limiting amino acids in protein malnourished areas, lactic fermentation of milk may be undesirable based on the chemical scores of the protein. However, the free amino acid content, including that of lysine and methionine, increases significantly upon lactic fermentation of milk. This predigestion of protein might result in an increased availability of the amino acids to offset the reduction in amino acids during fermentation and may even result in a better quality protein. Biological evaluation of the fermented milk proteins is necessary to evaluate their nutritional quality. L. bulgaricus was the most proteolytic of all the organisms used.     

喷雾干燥法制备双孢菇蛋白粉工艺优化及其营养价值评价

建立了喷雾干燥法制备双孢菇蛋白粉的工艺。以集粉率为指标,在单因素实验基础上,对进风温度、提取液蛋白质浓度、进料速率进行单因素及正交实验。结果表明:进风温度为165 ℃、提取液蛋白质浓度为40 mg/mL、进料速率为650 mL/h时,集粉率最高为38.8%。所得蛋白粉为奶白色,有鲜蘑菇香味。营养检测分析表明,双孢菇蛋白粉蛋白质含量为65.8%,碳水化合物为8.8%,粗脂肪为0.4%,粗纤维为3.8%,灰分为2.0%,水分为7.20%;氨基酸种类齐全,人体必需的8种氨基酸占氨基酸总量的41.0%;苏氨酸、异亮氨酸、赖氨酸、色氨酸四种必需氨基酸接近FAO/WHO标准。此外,还含有丰富的矿质元素,如重金属镉、铅、砷、汞含量均在国家规定的食品安全标准以内。