氨基酸微量元素螯合物在动物营养中的研究进展

从氨基酸微量元素螯合物的结构性质、生物利用率、营养特性、作用机理、制备、检测技术和生产应用现状等几个方面综述了它在动物营养中的最新研究进展，并就其存在问题和发展前景作了阐述。     

氨基酸锌化合物的研究进展

氨基酸锌作为一种重要的营养添加剂,在补充人体微量元素方面有着重要的作用,本文重点综述了氨基酸锌制备的方法以及制备过程中的影响因素。     

江西黄毛楤木氨基酸及微量元素的测定分析

用氨基酸分析仪、原子吸收分光光度计,对江西黄毛楤木中的营养成份进行分析研究。分别测定黄毛楤木根皮中氨基酸及微量元素含量。结果显示黄毛楤木根皮中含有16种氨基酸,总量为4.57mg·g-1,人体必需的8种氨基酸齐全,质量分数为1.71mg·g-1,占氨基酸总量的37.33％;含有多种人体必需的微量元素和宏量元素,尤其是zn,cu,K,Mg含量较其它种楤木要高。江西黄毛楤木富含氨基酸、微量元素,营养成分丰富,作为一种营养、保健、医药资源大有开发利用价值。     

云参化学成分研究Ⅱ.云参中氨基酸和微量元素的分析

本文分析测定了云参中的氨基酸和微量元素。结果表明。云参中含15种氨基酸,其中包括人体必须又只能由外界供给的7种氨基酸。同时,云参中还富含铁、硒、锌、锰等对人体具有重要营养、生理及临床意义的微量元素16种。     

海南野生石斛中必需微量元素与必需氨基酸的分析

本文对几种海南野生石斛(红头金石斛、重唇石斛、流苏金石斛、华石斛、密花石斛、刀叶石斛)及人工栽培的金钗石斛,进行了人体及植物必需微量元素和人体必需氨基酸的含量分析.结果表明:每种石斛中均含有植物和人体所需的7种微量元素和7种人体必需氨基酸,其中流苏金石斛的微量元素和氨基酸含量均高于其他种.人工栽培的金钗石斛其微量元素和氨基酸含量与海南野生石斛比较,差异不大,仅在Fe、Mn两个元素均低于野生种.本文还进行了植物必需微量元素与必需氨基酸相关性的讨论.     

口服水解蛋白及其营养合剂的制备与研究

本文通过以猪血为原料胰酶水解制备口服水解蛋白及营养合剂的研究,它含有多种必需氦基酸、微量元素、维生素,氨基酸同时促进各种离子吸收,动物对照实验表明该合剂能促进小鼠生长（Ｐ＜０．０１）。对创伤病人,病后、手术后的恢复健康结果令人满意。     

不同饲料添加剂对樱桃谷鸭生长性能及鸭舍有害气体含量的影响

选用1600羽出壳1 d的樱桃谷鸭随机分为8组,分别饲喂不同配比和组合的抗生素、沸石、微生态制剂(EM)、中草药和甘氨酸微量元素螯合物等添加剂饲料,探讨了不同饲料添加剂对肉鸭生长性能的影响,并对鸭舍的有害气体含量进行了研究。结果表明：饲料中添加沸石、EM、中草药对提高肉鸭的日增重和降低料肉比有显著效果(P＜0.05)。在1～14 d阶段,以混合添加沸石和EM对肉鸭体重、采食量和日增重的效果为最好(P＜0.05);添加抗生素、混合添加抗生素与沸石均能明显(P＜0.05)降低1～14 d樱桃谷鸭料肉比。在15～35 d阶段,添加沸石、EM、中草药均能提高肉鸭的饲料利用率,以混合添加沸石与中草药及甘氨酸微量元素螯合物效果最好,可显著(P＜0.05)提高肉鸭体质量、采食量和日增重,降低料肉比。添加沸石、EM、中草药均能降低鸭舍内NH₃、H₂S和CO₂的浓度,尤以添加沸石、甘氨酸微量元素螯合物、沸石与中草药混合物分别对降低NH₃、H₂S、CO₂浓度效果最好。说明沸石、中草药和EM可有效提高肉鸭生长性能,降低鸭舍内有害气体浓度。     

珍珠及珍珠层的化学成分比较研究

本文对不同产地及不同等级的淡水养殖褶纹冠蚌、三角帆蚌珍珠以及珍珠层中的氨基酸和微量元素进行了对比分析,结果表明,珍珠的来源不同,其氨基酸的种类及含量不同,各种微量元素的含量也有差异。研究表明,珍珠的等级与化学成分含量无相关性。     

酵母作为微量元素载体的研究及应用前景

微量元素是动物维持生命和发育的必需营养物质,它们的重要营养功能早已被人们所认识。微量元素的缺乏会造成人及动物营养不良并诱发多种疾病[1~2]。矿物质微量元素一般以无机盐形态补充,这种形式不利于动物吸收利用,有些微量元素在加工、储存及运输过程中不稳定易于氧化并与食品及饲料中其它成分协同配伍性差。近年来,不同来源的有机态微量元素逐渐被采用,大量试验表明,有机态微量元素较之无机态不仅利于动物吸收利用,而且某些有机微量元素,对增进动物健康,提高饲料转化率及生产性能,减少应激等方面均起很大作用[3~5]。螯合型氨基…     

中华鳖氨基酸和微量元素的分析与研究

本文对中华鳖的氨基酸、维生素、微量元素等营养物质进行了分析，并与其它十种水产品进行了比较，通过对中华鳖的化学分（ＣＳ）、氨基酸分（ＡＳ）和必需氨基酸指数（ＥＡＡＩ）的计算可知，中华鳖的微量元素构成较为合理，氨基酸构成与ＦＡＯ模式近似。     

Suppression of protein interactions by arginine: a proposed mechanism of the arginine effects

Arginine has been used to suppress protein aggregation and protein-protein or protein-surface interactions during protein refolding and purification. While its biotechnology applications are gradually expanding, the mechanism of these effects of arginine has not been fully elucidated. Arginine is more effective at higher concentrations, an indication of weak interactions with the proteins. The effects of weakly interacting additives, such as arginine, on protein solubility, stability and aggregation have been explained from three different approaches: i.e., (1) the effects of additives on the structure of water, (2) the interactions of additives with the amino acid side chains and peptide bonds and (3) the preferential interactions of additives with the proteins. Here we have examined these properties of arginine and compared with those of other additives, e.g., guanidine hydrochloride (GdnHCl) and certain amino acids and amines. GdnHCl is a strong salting-in agent and denatures proteins, while betaine is a protein stabilizer. Several amino acids and amine compounds, including betaine, which stabilize the proteins, are strongly excluded; i.e., the proteins are preferentially hydrated in these solutions. On the other hand, GdnHCl preferentially binds to the proteins. Arginine is intermediate between these two extreme cases and shows a more complicated pattern of interactions with the proteins. The effects of additives on water structure, e.g., the surface tension of aqueous solution of the additives and the solubility of amino acids in the presence of additives also shed light on the mechanism of the effects of the additives on protein aggregation. While arginine increases the surface tension of water, it favorably interacts with most amino acid side chains and the peptide bonds, a property shared with GdnHCl. Thus, we propose that while arginine is similar to GdnHCl in the amino acid level, arginine interacts with the proteins differently from GdnHCl.     

真菌菌丝体和发酵液的氨基酸分析比较

<正> 对云芝等7种真菌的菌丝体和发酵液分别测定17种氨基酸的含量,测定结果表明这些菌丝体中含有丰富的人体8种必需氨基酸,支链氨基酸及精氨酸,而芳香族氨基酸和含硫氨基酸却很低。7种真菌的菌丝体和它们的子实体,和三种食用蛋白(α-酪蛋白,卵白蛋白,大豆球蛋白)及4种粮食产品     

Why do solution additives suppress the heat‐induced inactivation of proteins? Inhibition of chemical modifications

Thermoinactivation of proteins is prevented by several kinds of solution additives such as chaotropes, amino acids, amino acid derivatives, and polyamines. Here, we investigated the molecular mechanisms of action of the various additives that prevent thermoinactivation of bovine pancreatic ribonuclease A and hen egg white lysozyme. The thermoinactivation of both proteins in the presence of additives showed clear correlations with deamidation and β-elimination of the proteins. Thus, experimental evidences indicated that the effects of additives on thermoinactivation of proteins are highly due to the suppression of chemical modifications. To our surprise, not only the suppressive effect of the additives on heat-induced inactivation but also that on the chemical modification of proteins is remarkably similar by comparison of two unrelated proteins. This finding indicates the generality of the effects of additives on heat-induced chemical modification of proteins.     

Oxidative refolding of rPA in l‐ArgHCl and in ionic liquids: A correlation between hydrophobicity,salt effects,and refolding yield

The ionic liquid 1‐ethyl‐3‐methyl imidazolium chloride (EMIM Cl) and the amino acid l‐ arginine hydrochloride (l ‐ArgHCl) have been successfully used to improve the yield of oxidative refolding for various proteins. However, the molecular mechanisms behind the actions of such solvent additives—especially of ionic liquids—are still not well understood. To analyze these mechanisms, we have determined the transfer free energies from water into ionic liquid solutions of proteinogenic amino acids and of diketopiperazine as peptide bond analogue. For EMIM Cl and 1‐ethyl‐3‐methyl imidazolium diethyl phosphate, which had a suppressive effect on protein refolding, as well as for l ‐ArgHCl favorable interactions with amino acid side chains, but no favorable interactions with the peptide backbone could be observed. A quantitative analysis of other ionic liquids together with their already published effects on protein refolding showed that only solvent additives within a certain range of hydrophobicity, chaotropicity and kosmotropicity were effective for the refolding of recombinant plasminogen activator. © 2014 Wiley Periodicals, Inc. Biopolymers 101: 1129–1140, 2014.     

Russian Article pp. 279-284

The accumulation of nucleic and protein (amino nitrogen) components as decomposition products of the process of autolysis of Saccharomyces cerevisiae yeast cells was studied at 50°C under the effect of various membranotropic additives. The influence of the added n-alcohols, n-fatty acids and several peptides was investigated in the range of concentration of 0.1–0.5 M. The maximal acceleration of the autolysis has been demonstrated under the effect of additives with a hydrophobicity of 7.5–8.5 ccal/M. In all the investigated concentrations stearic acid and octadecyl alcohol have an inhibitory influence. The role of the hydrophobic influences and the mechanism of autolysis are discussed.     

Mutations towards enantioselectivity adversely affect secretion of Pseudomonas aeruginosa lipase

Lipases are important biocatalysts used as detergent additives to manufacture biodiesel, and in particular, for the production of enantiopure compounds such as alcohols, amines and carboxylic acids. Extensive efforts were conducted trying to optimize lipase properties and lipase LipA of Pseudomonas aeruginosa comprises the best-studied example in terms of optimizing enantioselectivity by application of numerous directed evolution methods. Its enantioselectivity in the asymmetric hydrolysis of the model substrate 2-methyldecanoic acid p-nitrophenyl ester was increased from E=1.1 for the wild-type enzyme to E=51 for the best (S)-enantioselective variant which carried six amino acid exchanges. We have observed that overexpression of this variant in the homologous host resulted in only marginal yields of enzyme in the bacterial culture supernatant, suggesting that the enantioselective LipA variant was secreted with only low efficiency. Hence, we have analysed the secretion of this lipase variant and compared it to variants carrying either the respective single mutations or some combinations. We report here the identification of two amino acid substitutions located on the protein surface, which significantly impair lipase secretion.     

Characterization of protein fractions and amino acids in ensiled alfalfa treated with different chemical additives

Formic acid, formaldehyde, tannic acid or mixtures of two were studied on their effects on ensiled alfalfa (Medicago sativa L.) amino acids and N fractions by the Cornell Net Carbohydrate and Protein System (CNCPS). The alfalfa forage was a second cut and was wilted to a mean over-dry dry matter (DM) content of 330 g/kg. All silages were prepared as mini-silos using 100 ml polypropylene centrifuge tubes (50 g) on a small laboratory-scale, with the additives added in 20 ml aliquots/kg herbage fresh weight (FW). After 35 d of ensiling, most of forage true protein was converted to fraction A and all of the added additives reduced fraction A content in the ensiled forages (P<0.05). The content of fraction B₁ in all of the additive-treated silages was higher (P<0.05) than that in control silage. Large proportions of true protein in the tannic acid/formaldehyde- and formic acid/formaldehyde-treated silages were fractions B₂ and B₃, respectively. No difference was observed on fraction C content between the control silage and silages treated with additives except for the formaldehyde or tannic acid-treated silages. Amino acids were well preserved in additive-treated silages compared with the control silage. Concentration of total amino acid was higher in formic acid-treated silages than that in the control and the other additive-treated silages (P<0.05). The pattern of changes in individual amino acid in all of the silages indicated that branched chain amino acids and methionine were relatively well preserved during fermentation but the basic and acidic amino acids were not.     

Protein-free culture medium improvement: testing additives and their interactive effects in 96-well plates

In order to achieve a rational screening of additives suspected to improve antibody production over basal Dulbecco's Modified Eagle Medium (DMEM), we have developed a 96-well plate method for simultaneously testing individual and interactive effects of multiple additives. Cell viabilities were determined directly in each well by a colorimetric assay (MTT assay) and antibody production was measured by an enzyme-linked immunosorbent assay (ELISA) performed on well supernatants. Such as supplemented culture medium might considerably reduce production costs since commercial serum- or protein-free media are sold at serum-enriched basal medium costs. The CBM-P22 mouse cell line used in this study was shown to be sensitive to key amino acids, oxalacetic acid, ethanolamine and selenium at low cell density (<1 × 10⁵ cells·ml^–1). When these cells were inoculated in 96-well plates their antibody productivity was improved (sevenfold) by adding these additives to the basal DMEM as evidenced by ELISA absorbance readings. This improved productivity, obtained with the supplemented DMEM, named DOWSENs, is comparable to the one obtained with the commercial, but costly, protein-free hybridoma medium (PFHM II), taken here as the positive control. Results were confirmed by growing these cells in different media in standard (25 cm²) T-flask static culture. In addition, specific amino acid consumption, as analysed by HPLC, showed that asparagine and tryptophan when added to DMEM may improve antibody production, even if these amino acids are not limiting or highly consumed in PFHM II. Using this 96-well plate assay allows the assessment of a large number of different assays with a few milligrams of the product(s) tested. This is a rapid technique that gives results for additives that are not easily quantified by analytical techniques or for the study of interactive effects, which is time consuming and labour-intensive when done in individual T-flasks. Correspondence to: J. Côté     

Suppression of epimerization by cupric (II) salts in peptide synthesis using free amino acids as amino components.

An epimerization-free system for coupling N-protected peptides with free amino acids was developed. A number of inorganic substances were tested as epimerization suppressant additives during the coupling by various methods (carbodiimide plus additives, uronium salts, Woodward's reagent-K, isobutyl-chloroformate, etc.). Some of them (ZnCl2, RbClO4, LiCl, SnCl4, AlCl3, etc.) in combination with some coupling methods can guarantee coupling with minimal epimerization (D-epimer < 1%). But only a simultaneous use of 1-hydroxybenzotriazole and Cu2+ ions as additives in carbodiimide-mediated peptide couplings appeared to give a standard result (D-epimer < 0.1%). There was no epimerization even in the case when N-methyl amino acid (sarcosine) was used as an amino component, while in the absence of Cu2+ ions an unacceptable level of epimerization was observed (D-epimer, 22% for carbodiimide with the 1-hydroxybenzotriazole method). So far it has been considered that Cu2+ ions prevent obtaining peptides in high yields (< 90%) by various coupling methods. We have found that the use of 1-hydroxybenzotriazole, CuCl2 and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide instead of dicyclohexylcarbodiimide provides a possible method for obtaining the desired peptides in 90-99% yields without epimerization. All these results were shown by employing several model peptide couplings with free amino acids as amino components dissolved in an effective solvent system which readily dissolved them.     

The Effect of Nisin and Monensin on Ruminal Fermentations In Vitro

When mixed ruminal bacteria and alfalfa were incubated in vitro, monensin and nisin both inhibited methane production so long as the concentrations were greater than 1 μM. Monensin- and nisin-dependent methane depressions caused a decrease in the acetate to propionate ratio (4.5 to 3.0). Total volatile fatty acid production was decreased by both monensin and nisin addition at concentrations greater than 2 μM. Starch-digesting ruminal bacteria were initially inhibited by monensin and nisin, but this effect disappeared after two to four transfers. Nisin always inhibited cellulolytic bacteria, but the nisin-dependent inhibition of cellulose digestion was no greater than the inhibition caused by monensin. Monensin and nisin also inhibited amino acid degradation, and nisin was more effective than monensin in controlling the growth of Clostridium aminophilum, an obligate amino acid-fermenting ruminal bacterium that can tolerate low concentrations of monensin. Because nisin was as potent as monensin, bacteriocins such as nisin may have potential as feed additives. Received: 2 December 1996 / Accepted: 10 February 1997