花粉破壁法对花粉蜂蜜酒中氨基酸含量的影响

以蜂蜜为主料,加入花粉,经过调配、发酵、过滤、陈酿后获得氨基酸含量丰富的酒精饮料。花粉破壁分别采用超声破碎法、酶法和温差破壁法这几种不同方式处理,并添加于蜂蜜发酵液中,经酒精发酵,制得富含活性成分的花粉蜂蜜酒。最后利用毛细管电泳法来测定成品酒中的氨基酸,通过分析优化出最佳的花粉破壁方法。研究结果表明花粉经过温差破壁法处理后加入发酵液中,其成品的氨基酸含量最高。该产品不仅营养价值高,风味独特,而且还具有良好的保健作用。     

Kinetic resolution of amino acid esters catalyzed by lipases

Racemic amino acids were resolved by lipase via hydrolysis of their esters. Lipases (Pseudomonas lipase from Amano PS, Rhizopus lipase from Serva, and porcine pancrease lipase from Sigma) could selectively hydrolyze the L-amino acid esters in aqueous solution with high reactivities and selectivities. The effect of the structural changes in the ester moiety on the stereoselectivity of the lipases was also investigated using D ,L -homophenylalanine as a model. Procedures were developed for the resolution of natural and unnatural amino acids. © 1996 Wiley-Liss, Inc.     

Comparative study of the free amino acid compositions and contents in three different botanical origins of Coptis herb

Free amino acids are important bioactive ingredients in herbs and herbal products. Coptis herbs contain a variety of free amino acids; however, studies have not yet analyzed the relationship between free amino acids and species of Coptis herbs. In the current study, the contents of 20 free amino acids in Coptis chinensis Franch., Coptis teeta Wall., and Coptis deltoidea C.Y. Cheng et Hsiao were determined using an automatic amino acid analyzer to evaluate the differences between the three species. We found that the major amino acids (Asn, Arg, and γ-aminobutyric acid (GABA)) were significantly more in C. chinensis than in the other two species. In addition, Asp content was significantly more in C. deltoidea. The study concluded that Asn can be used to identify C. deltoidea, C. chinensis, and C. teeta, and Gln and Arg can be used to distinguish C. teeta from C. chinensis and C. deltoidea. These findings suggested that free amino acids as active ingredients can be used to identify Coptis herb species.     

Regulation of the plasma amino acid profile by leucine via the system L amino acid transporter

Plasma concentrations of amino acids reflect the intracellular amino acid pool in mammals. However, the regulatory mechanism requires clarification. In this study, we examined the effect of leucine administration on plasma amino acid profiles in mice with and without the treatment of 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH) or rapamycin as an inhibitor of system L or mammalian target of rapamycin complex 1, respectively. The elevation of plasma leucine concentration after leucine administration was associated with a significant decrease in the plasma concentrations of isoleucine, valine, methionine, phenylalanine, and tyrosine; BCH treatment almost completely blocked the leucine-induced decrease in plasma amino acid concentrations. Rapamycin treatment had much less effects on the actions of leucine than BCH treatment. These results suggest that leucine regulates the plasma concentrations of branched-chain amino acids, methionine, phenylalanine, and tyrosine, and that system L amino acid transporters are involved in the leucine action.     

Effects of total gastrectomy on plasma silicon and amino acid concentrations in men

The aim of the study was to determine one-year effects of total gastrectomy on plasma silicon and free amino acid concentrations in patients and evaluate changes of volumetric bone mineral density (vBMD) in lumbar spine. Eight patients were enrolled to the control (CTR) group. Six patients subjected to total gastrectomy (GX group) were included to the experimental group. vBMD in trabecular and cortical bone was measured in lumbar vertebrae at baseline (before surgery) and one year later using quantitative computed tomography. Plasma concentrations of silicon and free amino acids were determined at baseline and one year later using photometric method and ion-exchange chromatography. Body weights within CTR and GX groups were not different after one-year follow-up when compared to the baseline values (P > 0.05). An average annual decrease of vBMD in the trabecular bone in the gastrectomized patients reached 15.0% in lumbar spine and was significantly different in comparison to the percentage changes observed in CTR group (P = 0.02). One-year percentage change of vBMD in the cortical bone in L₁ and L₂ has shown significantly decreased values by 10.5 and 9.1% in the GX group when compared to the percentage change observed in the controls (P < 0.05). Plasma concentration of adipic acid was significantly higher by 101.6% one year after total gastrectomy procedure in the patients when compared to the baseline value (P = 0.01). Plasma concentration of silicon was significantly lowered by 26.7% one year after the total gastrectomy when compared to the baseline value (P = 0.009). Total gastrectomy in patients has induced severe osteoporotic changes in lumbar spine within one-year period. The observed osteoporotic changes were associated with decreased plasma concentration of silicon indicating importance of exocrine and endocrine functions of stomach for silicon homeostasis maintenance. Gastrectomy-induced bone loss was not related to decreased amino acid concentration in plasma obtained from overnight fasted patients.     

兴安落叶松游离氨基酸的动态变化与损伤因子的关系

为研究不同损伤因子及其不同损伤程度对落叶松游离氨基酸组分及其含量的影响,用剪叶和落叶松毛虫(Dendrolimussuperans)取食损伤兴安落叶松(Larixgmelinii)幼苗,使用HPLC柱前AccQ.Tag衍生法测定针叶内游离氨基酸的组成及含量的变化。结果表明,松针中含有17种游离氨基酸,以苯丙氨酸含量最高,损伤15d后,剪叶4枝比对照增加36.6%,虫害4枝增加的百分率达到98.3%。剪叶4枝15d后及虫害4枝5d后,损伤25%与50%之间氨基酸总含量差异显著(P0.05),且昆虫取食4枝25%、50%、75%后松针内游离氨基酸总含量的变化趋势与剪叶处理后总氨基酸含量的变化一致,均在处理10d后呈现较高的含量,虽虫害后的氨基酸总含量高于剪叶损伤后的总含量,但二者之间差异不显著。剪叶4枝、8枝、12枝3个损伤程度后针叶内的游离氨基酸含量无显著差异,说明剪叶4枝即能超过落叶松的耐受阈值,达到防御诱导效应。两种处理下多种游离氨基酸的含量均有下降,其中8种氨基酸含量存在显著差异(P0.05),即丝氨酸、谷氨酸、甘氨酸、组氨酸、脯氨酸、半胱氨酸、赖氨酸和苯丙氨酸。表明人工模拟剪叶的损伤方式与虫害都可诱导松树氨基酸组分及含量的变化,而间接影响害虫的生长发育。     

Effects of long-term antibiotic treatment on mice urinary aromatic amino acid profiles

The gut microbiota–host co-metabolites are good indicators for representing the cross-talk between host and gut microbiota in a bi-direct manner. There is increasing evidence that levels of aromatic amino acids (AAAs) are associated with the alteration of intestinal microbial community though the effects of long-term microbial disturbance remain unclear. Here we monitored the gut microbiota composition and host–microbiota co-metabolites AAA profiles of mice after gentamicin and ceftriaxone treatments for nearly 4 months since their weaning to reveal the relationship between host and microbiome in long- term microbial disturbances. The study was performed employing targeted LC-MS measurement of AAA-related metabolites and 16S RNA sequence of mice cecal contents. The results showed obvious decreased gut microbial diversity and decreased Firmicutes/Bacteroidetes ratio in the cecal contents after long-term antibiotics treatment. The accumulated AAA (tyrosine, phenylalanine and tryptophan) and re-distribution of their downstreaming metabolites that produced under the existence of intestinal flora were found in mice treated with antibiotics for 4 months. Our results suggested that the long-term antibiotic treatment significantly changed the composition of the gut microbiota and destroyed the homeostasis in the intestinal metabolism. And the urinary AAA could be an indicator for exploring interactions between host and gut microbiota.     

The effect of caffeine on some mouse brain free amino acid levels

Changes in free amino acids were examined in the central nervous system of mice treated with caffeine for three weeks. Caffeine was administered in the drinking water, and at the end of three weeks the level of caffeine in the cerebral cortex was 113±19 g/g. When amino acid levels in cerebral hemispheres, midbrain, pons and medulla, and cerebellum were measured a significant increase in glutamine levels was found in all four regions. Glycine, alanine, serine, threonine, and GABA were significantly reduced in some regions. Caffeine appears to alter some of the metabolic or transport processes regulating amino acid pools in the brain. The decrease of GABA found in pons and medulla may contribute to the observed increase in reflex excitability after caffeine.Special issue dedicated to Dr. Elling Kvamme     

Free amino acid concentrations in milk: Effects of microwaveversus conventional heating

Summary Microwave effects on free amino acid concentrations in milkversus a water bath heating were investigated in view of their importance for infant growth. Concentrations of few amino acids, such as aspartate, serine or lysine, are unchanged whatever the way and the temperature of heating. In contrast, tryptophan concentrations decreased similarly whatever the way of heating (110 ± 3µmol/l before heatingvs 84 ± 4µmol/l after 30°C microwave heating, p < 0.05). On the contrary, concentrations of glutamate and glycine increased more after water bath heating at 90°C (325 ± 4 and 101 ± 1µmol/l, respectively) than after microwave heating (312 ± 4 and 95 ± 1µmol/l, respectively, p < 0.05) suggesting milk proteolysis. Moreover, the accumulation of ammonia observed at 90°C with the water bath together with increase Glu levels might reflect a degradation of glutamine. An ornithine enrichment, more evident with microwave heating, was shown and could be of interest as it is a polyamine precursor. Also, considering few variations of free amino acid concentrations and the time saved, microwave heating appears to be an appropriate method to heat milk.     

Recognition of specific patterns of amino acid inhibition of growth in higher plants, uncomplicated by glutamine-reversible `general amino acid inhibition''

The complexity of the regulatory mechanisms that govern amino acid biosynthesis, particularly in multibranched pathways, frequently results in sensitivity to growth inhibition by exogenous amino acids. Usually the inhibition caused by a given amino acid(s) is relieved by another amino acid(s), thus indicating the cause of inhibition to be a specific interference with endogenous formation of the latter amino acid(s). We recently summarized the evidence that Nicotiana silvestris (and probably most higher plants), in suspension culture, exhibits a separate phenomenon of amino acid mediated growth inhibition called general amino acid inhibition. Every amino acid provokes general amino acid inhibition except for

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-glutamine. In fact,

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-glutamine completely overcomes general amino acid inhibition. We have now demonstrated that specific amino acid inhibition can be recognized and characterized at the level of growth inhibition without interference caused by general amino acid inhibition by the simple provision of exogenous

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-glutamine. Several examples of specific amino acid inhibition of growth were demonstrated in N. silvestris. In one case,

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-threonine inhibits growth partially in the presence of

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-glutamine. The residual amino acid inhibition was overcome by the additional presence of

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-lysine and

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-methionine, indicating that exogenous

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-threonine specifically inhibits the biosynthesis of both

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-lysine and

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-methionine. As a second example, the

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-valine-mediated inhibition of growth that persisted in the presence of

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-glutamine was overcome by

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-isoleucine, indicating that exogenous

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-valine inhibits

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-isoleucine biosynthesis. The use of amino acid analogs as experimental tools for biochemical-genetic studies in higher plants is also complicated by general amino acid inhibition. Conditions were demonstrated under which p-fluorophenylalanine and m-fluorotyrosine could be used as specific antimetabolites of

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-phenylalanine and

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-tyrosine biosynthesis without interference from general amino acid inhibition. We thus present a rigorous basis for recognition of specific relationships between metabolic branches that can guide detailed enzymological analyses.     

Endogenous excitatory amino acid release from brain slices and astrocyte cultures evoked by trimethyltin and other neurotoxic agents

Trimethyltin (TMT) is a toxic alkyltin compound that is known to produce neuronal necrosis in the CNS. The present study examined the effects of TMT on the release of excitatory amino acids (EAA) from cortical slices prepared from adult and aged (24 months old) rats. The calcium dependence of TMT-induced EAA efflux was evaluated and compared to other neurotoxic agents. The actions of TMT were also evaluated in an astrocyte culture model to assess glial contributions to TMT-induced EAA efflux. TMT (10–1000 M) evoked a dose-related increase in GLU and ASP efflux during a 30 min incubation period and this efflux was sustained or slightly higher during a 15 min recovery period. TMT-stimulated GLU efflux was not altered in aged rats. TMT-induced GLU efflux was significantly reduced by removing extracellular calcium and including 10 M EGTA in the incubation media. Calcium channel blockers (nifedipine, verapamil, flunarizine, amiloride, neomycin) and MK-801 did not significantly attenuate TMT-induced GLU efflux. Diltiazem (25 M) produced modest but inconsistent reductions in TMT-induced GLU efflux from brain slices, and significantly inhibited the leakage of lactate dehydrogenase (LDH) from TMT-treated astrocyte cultures. TMT did not increase GLU efflux from glial cultures during a 30 min incubation period, but did significantly elevate GLU efflux during the 15 min recovery period. TMT evoked the release of EAA by both calcium dependent and independent mechanisms in brain slices. TMT at high concentrations also produced a delayed increase in glial GLU efflux. These studies suggest that excitotoxic mechanisms may contribute to TMT-induced neurotoxicity.     

Effects of branched-chain amino acids on plasma amino acids in amyotrophic lateral sclerosis

Summary Although the cause of amyotrophic lateral sclerosis (ALS) remains unknown, biological findings suggest that the excitatory amino acid glutamate contributes to the pathogenesis of ALS. In previous studies of ALS, the therapeutic effect of the branched-chain amino acids (BCAAs) leucine, valine and isoleucine has been evaluated. The present study aimed at investigating the acute effect of BCAAs on plasma glutamate levels in ALS patients. Following two oral doses of BCAAs, significantly increased plasma levels were seen for valine (500%), isoleucine (1,377%) and leucine (927%), however the plasma level of glutamate was not affected. The plasma level of several other amino acids (tryptophan, tyrosine, phenylalanine and methionine) were found decreased after oral BCAAs, which may indicate a diminution in the rate of degradation of muscle protein and/or an increase in tissue disposal of amino acids.     

Nature of amino acid substitutions in homologous proteins during evolution

Replacement substitutions of mitochondrial cytochrome $\text{c}$ and α- and β-chains of haemoglobin have been studied by considering the structural similarity among amino acid residues at the secondary and tertiary structural levels. Secondary structural similarity explains ～70% while tertiary structural similarity explains ～50% of observed replacements for most of the cases. These structural similarities could not account for all the replacement substitutions. The study was extended to consider the composition of codons, and the chemical nature and polarity of the replacing and replaced residues. These also could not individually account for all the affected replacements. In general, no property of amino acid residues is conserved for substitutions occurring at any single position during evolution of proteins.     

Porcine pancreatic lipase-catalized enantioselective hydrolysis of N-protected amino acid methyl-esters

Summary A preparative-scale enantioselective hydrolysis of racemic methyl esters of several N-protected amino acid has been carried out by using crude porcine pancreatic lipase (Triacylglycerol lipase, EC 3.1.1.3) PPL as a hydrolytic enzyme. In all cases 50% of the racemic methyl ester was hydrolysed to the N-protected L-amino acid with high yield and high optical purity.Hydrolysis rates were very close related not only to the amino acid structure but also to the steric and/or electronic nature of the ester and N-protecting groups. Thus, the very convenient ester methyl group can be enantioselectively hydrolysed with PPL when N-protecting group is a carbonyl derivative, as it is the usual benzoyl group.     

Aspects of astrocyte energy metabolism,amino acid neurotransmitter homoeostasis and metabolic compartmentation

Astrocytes are key players in brain function; they are intimately involved in neuronal signalling processes and their metabolism is tightly coupled to that of neurons. In the present review, we will be concerned with a discussion of aspects of astrocyte metabolism, including energy-generating pathways and amino acid homoeostasis. A discussion of the impact that uptake of neurotransmitter glutamate may have on these pathways is included along with a section on metabolic compartmentation.     

Metabolic memory effect of the saturated fatty acid, palmitate, in monocytes

Hyperglycaemia has a deferred detrimental effect on glucose metabolism, termed “metabolic memory”. Elevated saturated fatty acids promote insulin resistance, hyperglycaemia and associated atherosclerotic complications, but their effect on “metabolic memory” is unknown. Therefore we investigated whether basal and insulin-stimulated (10⁻⁶ M for 12 h) glucose (2-deoxy-d-[³H]-glucose) uptake was affected by palmitate pre-treatment human THP-1 monocytes. Palmitate-induced a time-dependent and concentration-dependent inhibition of insulin-stimulated glucose uptake, showing almost complete abolition of the insulin-stimulatory effect with 300 μM palmitate. Basal glucose uptake was unaffected by palmitate. When palmitate was washed out, the inhibitory effect on insulin-stimulated glucose uptake persisted for at least 60 h.     

Carbohydrate and free amino acid contents in tomato plants grown in media with bicarbonate and nitrate or ammonium

Tomato plants were cultivated (from 2 to 23 days after germination) in media with NO ₃ ⁻ , NH ₄ ⁺ , or a mixture of both forms in different proportions used as the N source given with or without 5 mol dm⁻³ HCO ₃ ⁻ . The accumulation of soluble sugars (reducing sugars and sucrose) and free amino acids was higher in the roots and leaves of NH ₄ ⁺ -fed plants than in NO ₃ ⁻ -fed plants. Starch accumulation in NH ₄ ⁺ -fed plants was higher in leaves (about 28%) and lower in roots (about 37%) in comparison with that of NO ₃ ⁻ -fed plants. Plants cultivated in media containing a mixture of NO ₃ ⁻ /NH ₄ ⁺ were characterized by a lower content of sugars and amino acids accumulation in comparison with that in plants fed with NO ₃ ⁻ or NH ₄ ⁺ . An elevated HCO ₃ ⁻ concentration in the rhizosphere stimulated the accumulation of soluble sugars and free amino acids in all the experimental variants. There were only small differences in the starch content.     

Hormonal regulation of hepatic amino acid transport

The transport of 2-aminoisobutyric acid (AIB) into liver tissue was increased by both insulin and glucagon. We have now shown that these hormones do not stimulate the same transport system. Glucagon, possibly via cAMP, increased the hepatic uptake of AIB by a mechanism which resembled system A. This glucagon-sensitive system could be monitored by the use of the model amino acid MeAIB. In contrast, the insulin-stimulated system exhibited little or no affinity for MeAIB and will be referred to as system B. On the basis of other reports that the hepatic transport of AIB is almost entirely Na⁺ dependent and the present finding that the uptake of 2-aminobicyclo [2,2,1] heptane-2-carboxylic acid (BCH) was not stimulated by either hormone, we conclude that system B is Na⁺ dependent. Furthermore, insulin added to the perfusate of livers from glucagon-pretreated donors suppressed the increase in AIB or MeAIB uptake. Depending upon the specificities of systems A and B, both of which are unknown for liver tissue, the insulin/glucagon ratio may alter the composition of the intracellular pool of amino acids.     

Effects of l-leucine in ovo feeding on thermotolerance,growth and amino acid metabolism under heat stress in broilers

Recently, we found that in ovo feeding of l-leucine (l-Leu) afforded thermotolerance, stimulated lipid metabolism and modified amino acid metabolism in male broiler chicks. However, the effects of in ovo feeding of l-Leu on thermoregulation and growth performance until marketing age of broilers are still unknown. In this study, we investigated the effects of in ovo feeding of l-Leu on body weight (BW) gain under control thermoneutral temperature or chronic heat stress. We measured changes of body temperature and food intake, organ weight, as well as amino acid metabolism and plasma metabolites under acute and chronic heat stress in broilers. A total of 168 fertilized Chunky broiler eggs were randomly divided into 2 treatment groups in experiments. The eggs were in ovo fed with l-Leu (34.5 µmol/500 µl per egg) or sterile water (500 µl/egg) during incubation. After hatching, male broilers were selected and assigned seven to nine replicates (one bird/replicate) in each group for heat challenge experiments. Broilers (29- or 30-day-old) were exposed to acute heat stress (30 ± 1°C) for 120 min or a chronic heat cyclic and continued heat stress (over 30 ± 1°C; ages, 15 to 44 days). In ovo feeding of l-Leu caused a significant suppression of enhanced body temperature without affecting food intake, plasma triacylglycerol, non-esterified fatty acids, ketone bodies, glucose, lactic acid or thyroid hormones under acute heat stress. Daily body temperature was significantly increased by l-Leu in ovo feeding under chronic heat stress. Interestingly, in ovo feeding of l-Leu caused a significantly higher daily BW gain compared with that of the control group under chronic heat stress. Moreover, some essential amino acids, including Leu and isoleucine, were significantly increased in the liver and decreased in the plasma by l-Leu in ovo feeding under acute heat stress. These results suggested that l-Leu in ovo feeding afforded thermotolerance to broilers under acute heat stress mainly through changing amino acid metabolism until marketing age.     

The amino acid composition of tissue protein is affected by the total sulfur amino acid supply in growing pigs

The factorial approach to assess the amino acid (AA) requirements of pigs is based on the assumption that the AA composition of body protein is constant. However, there are indications that this assumption may not be valid because the AA composition of body protein can be affected by the AA supply. The extent to which different tissues are affected by an AA deficiency is unknown. The objective of this study was to investigate the effect of feeding pig diets with a deficient or sufficient total sulfur AA supply (TSAA; Met+Cys) from 6 to 23 weeks of age on tissue composition and meat quality. The deficient diet (TSAA–) provided 24% Met : Lys and 51% TSAA : Lys on a standardized ileal digestible basis, which are 19% and 16% below the recommended requirements, respectively. The sufficient diet (TSAA+) provided 33% Met : Lys and 60% TSAA : Lys. Diets were offered slightly below the ad libitum feed intake capacity of the pigs. Pigs offered diet TSAA– had a lower average daily gain, lower weights of the longissimus dorsi (LM) and rhomboideus muscles (RM), and of selected skin sections (P<0.05). The weight of different sections of the small intestine and the liver was not affected by the diet. The protein content of the LM and RM decreased in pigs offered diet TSAA– (P<0.05), whereas the protein content of other tissues was not affected. The TSAA supply affected the AA composition (g/16 g N) of protein in all tissues, but the Met content was changed only in the liver (P<0.05). Pigs receiving diet TSAA– had a lower Cys content in the RM and in the distal jejunum and ileum (P<0.01). The deficient TSAA supply resulted in a lower carcass weight and higher muscle glycogen stores (P<0.05), but did not affect other meat quality traits. The results of this study indicate that the muscles, jejunum and ileum respond more to a prolonged AA deficiency than the liver. In addition, the observed changes in AA composition of tissue protein question the use of a constant AA profile of retained protein to assess AA requirements.