Metagenomic analysis revealing the metabolic role of microbial communities in the free amino acid biosynthesis of Monascus rice vinegar during fermentation

Free amino acid(FAA)is the important component of vinegar that infl uences quality perception and consumer acceptance.FAA is one of the major metabolites produced by microorganisms;however,the microbial metabolic network on FAA biosynthesis remains unclear.Through metagenomic analysis,this work aimed to elucidate the roles of microbes in FAA biosynthesis during Monascus rice vinegar fermentation.Taxonomic profiles from functional analyses showed 14 dominant genera with high contributions to the metabolism pathways.The metabolic network for FAA biosynthesis was then constructed,and the microbial distribution in different metabolic pathways was illuminated.The results revealed that 5 functional genera were closely involved in FAA biosynthesis.This study illuminated the metabolic roles of microorganisms in FAA biosynthesis and provided crucial insights into the functional attributes of microbiota in vinegar fermentation.

Effect of borneol, moschus, storax, and acorus tatarinowii on expression levels of four amino acid neurotransmitters in the rat corpus striatum

The present study collected cerebrospinal fluid samples from the corpus striatum in rats treated with borneol, moschus, storax, and acorus tatarinowii using brain microdialysis technology. Levels of excitatory neurotransmitters aspartic acid and glutamate, as well as inhibitory neurotransmitters glycine and ^-aminobutyric acid, were measured in samples using reversed-phase high-performance liquid chromatography, phosphate gradient elution, and fluorescence detection. Results showed that concentrations of all four amino acid neurotransmitters significantly increased in the corpus striatum following treatment with borneol or moschus, but effects due to borneol were more significant than moschus. Acorus tatarinowii treatment increased ^-aminobutyric acid expression, but decreased glutamate concentrations. Storax increased aspartic acid concentrations and decreased glycine expression. Results demonstrated that borneol and moschus exhibited significant effects on con amino acid neurotransmitter expression; storax exhibited excitatory effects and acorus tatarinowii resulted in inhibitory effects.

Effects of Subchronic Aluminum Exposure on Amino Acids Neurotransmitters in Chicken Brain

To investigate the effects of aluminium （Al） exposure on amino acid neurotransmitters, the chickens with different levels of subchronic Al poisoning were estabolished by continuous peritoneal injection of fixed volume and different concentrations of gradient of aluminium trichloride （AlCl3）. The levels of amino acid neurotransmitters in chicken brains were determined by high performance liquid chromatography （HPLC） after being exposed of Al for 60 days, and Al levels in serum and brain tissue were determined by atomic absorption spectrophotometry （AAS）. The results showed that Glu levels increased with the increase of Al, but there was no significant difference compared with the control. The levels of Al, Asp, Gly, GABA and Tau were significantly higher in Al-treated groups than those in the control. The results indicated that Al intoxication led to excitatory neurotoxicity.

Analysis of Amino Acids in a Single Human Red Blood Cell by Capillary Zone Electrophoresis with Intracellular NDA- derivatization and Electrochemical Detection

A novel method for determination of amino acids in individual human red blood cells has been developed. In this method, the derivatization reagents (NDA and CN-) are introduced into living cells by electroporation. After completion of derivatization, the amino acids in a single cell is determined by capillary zone electrophoresis with end-column amperometric detection.

Meta-analysis of soybean amino acid QTLs and candidate gene mining

The composition and quantity of amino acids influence the protein content and nutritional value of soybeans and also have an important impact upon soybean quality. After integrating and proofreading 140 original QTLs associated with amino acid contentfrom soybase（http：//www.soybase.org/）, 138 QTLs were further analyzed to determine high-confidence QTL regions. Meta-analysis was first carried out using the Bio Mercator ver. 2.1 software, yielding 33 consensus QTLs. The consensus QTL confidence intervals（CIs） ranged from 0.07 to 19.85 Mb. Next, the overview method was used to optimize the CIs, and 57 ＂real＂ QTLs were mapped. Candidate genes in the consensus QTL regions were obtained from Phytozome and were annotated using the Gene Ontology（GO）, Kyoto Encyclopedia of Genes and Genomes（KEGG）, Swissprot, and gene annotation databases. Finally, 16 unpublished candidate genes controlling the content of five types of amino acids were identified with Blast. These results laid the foundation for fine mapping of soybean amino acid-related QTLs and marker-assisted selection.

The structural analysis of protein sequences based on the quasi-amino acids code

Proteomics is the study of proteins and their interactions in a cell. With the successful completion of the Human Cenome Project, it comes the postgenome era when the proteomics technology is emerging. This paper studies protein molecule from the algebraic point of view. The algebraic system （∑, ＋, ＊） is introduced, where ∑ is the set of 64 codons. According to the characteristics of （∑, ＋, ＊）, a novel quasi-amino acids code classification method is introduced and the corresponding algebraic operation table over the set ZU of the 16 kinds of quasi-amino acids is established. The internal relation is revealed about quasi-amino acids. The results show that there exist some very close correlations between the properties of the quasi-amino acids and the codon. All these correlation relationships may play an important part in establishing the logic relationship between codons and the quasi-amino acids during the course of life origination. According to Ma F et al （2003 J. Anhui Agricultural University 30 439）, the corresponding relation and the excellent properties about amino acids code are very difficult to observe. The present paper shows that （ZU, ＋,×） is a field. Furthermore, the operational results display that the eodon tga has different property from other stop codons. In fact, in the mitochondrion from human and ox genomic codon, tga is just tryptophane, is not the stop codon like in other genetic code, it is the case of the Chen W C et al （2002 Acta Biophysiea Siniea 18（1） 87）. The present theory avoids some inexplicable events of the 20 kinds of amino acids code, in other words it solves the problem of ＇the 64 codon assignments of mRNA to amino acids is probably completely wrong＇ proposed by Yang （2006 Progress in Modern Biomedicine 6 3）.

Analysis of amino acid constituents of gallstones

IM To seek effective drugs to dissolve bilirubin, glycoprotein and black stones and better lithotriptics for cholesterol stones, and study the amino acid constituents of gallstones.METHODS According to the characteristics of infrared spectrum and the contents of bilirubin tested by semiquantitative chemical analysis, 30 of 148 cases of gallstones were selected and divided into 5 groups. Amino acids of the 30 cases were detected by high speed chromatography.RESULTS The quantity of amino acids in black stones was the highest (2269mg/g) and that of pure cholesterol stones was the lowest, only 14mg/g in a case. In five groups of gallstones, the contents of amino acids were: black stone > mixed bilirubin stone and glucoprotein stone > mixed cholesterol stone > pure cholesterol stone. The proportions were: 9595∶2902 and 2805∶578∶1. Aliphatic amino acids accounted for about 50% of the total amino acids in gallstones and glycine 153% of the total amount of 17 kinds of amino acids.CONCLUSION In mixed stones, the higher level of bilirubin, the higher contents of amino acids. The acidic amino acids of bilirubin stones were relatively higher than those of cholesterol stones.

Free amino acid (FAA) analysis of phloem sap in hybrid rice Shanyou 63 and its parents

Hyhrid rice has greatly contributed to rice production inChina. But concurrently a dramatic upsurge of rice plant-hopper occurred, particularly for the whitebacked plant-hopper (WBPH), Sogatella furcifera. It has become animportant economic insect pest of hybrid rice, although ithad only been a minor herbivore of rice before nation-wideexploitation of hybrid rice.

Comparative Analysis of Nutrient Composition of Caulerpa lentillifera from Different Regions

This study investigated the difference of nutrient composition in Caulerpa lentillifera collected from different regions.The nutrient compositions of C.lentillifera from China's Hainan and Shandong provinces were determined and compared with those from Semporna(Malaysia),Sabah(Malaysia),Petchburi(Thailand),and two other species of seaweeds(Malaysia).The results showed that the polysaccharide and protein contents of C.lentillifera from Hainan(44.82%±0.98%and 12.5%±0.70%),Shandong(43.22%±1.42%and 14.7%±0.72%),Petchburi(59.27%±2.31%and 12.49%±0.30%),and Sabah(44.02%±2.01%and 19.38%±1.48%)were higher than those of Eucheuma cottonii(26.49%±3.01%and 9.76%±1.33%)and Sargassum polycystum(33.49%±1.70%and 5.40%±0.07%),respectively.The polyunsaturated fatty acid contents in C.lentillifera from Hainan(29.98%)and Shandong(22.11%)were higher than that in Semporna(16.76%±0.27%),Sabah(9.49%),and S.polycystum(20.34%±0.43%),but lower than that in E.cottonii(51.55%±0.57%).In Caulerpa lentillifera of Hainan and Shandong provinces,the essential amino acid(EAA)/total amino acid(TAA)ratios were 35.22%and 38.78%,respectively;and the EAA/ETAA ratios were 54.36%and 63.35%,respectively.The EAA composition of C.lentillifera was close to the ideal model of Food and Agriculture Organization/World Health Organization.C.lentillifera was rich in mineral elements,including calcium((3315.85±127.55)mg(100 g)^-1 and(3728.35±92.38)mg(100 g)^-1),magnesium((6715.74±82.58)mg(100 g)^-1 and(8128.59±242.72)mg(100 g)^-1),and trace elements,including iron((510.65±5.47)mg(100 g)^-1 and(1972.97±183.35)mg(100 g)^-1),selenium((1.04±0.08)mg(100 g)^-1 and(0.83±0.15)mg(100 g)^-1),and zinc((33.90±0.13)mg(100 g)^-1 and(11.75±2.05)mg(100 g)^-1).These results showed that C.lentillifera was more nutritious than S.polycystum or E.cottonii in terms of polysaccharide,protein,and fatty acid contents.Furthermore,both C.lentillifera species collected from Hainan and Shandong provinces show more basic nutrients.Therefore,C.lentillifera has important development and application prospects.

Evaluation of the Protein Requirement in Chinese Young Adults Using the Indicator Amino Acid Oxidation Technique

Objective To accurately calculate the protein requirements in Chinese young adults using the indicator amino acid oxidation technique. Methods Nine women and ten men received a restricted daily level of protein intake （0.75, 0.82, 0.89, 0.97, and 1.05 g/kg）, along with L-[1-13C]-Ieucine. Subjects＇ protein requirement was determined by a biphasic linear regression crossover analysis of F13C02 data. In doing so, a breakpoint at the minimal rate of appearance of 13C02 expiration specific to each level of dietary protein was identified. This trial was registered with the Chinese clinical trial registry as ChiCTR-ONC-11001407. Results The Estimated Average Requirement （EAR） and the Recommended Nutrient Intake （RNI） of protein for healthy Chinese young adults were determined to be 0.87 and 0.98 g/（kg.d）, respectively, based on the indicator amino acid oxidation technique. Conclusion The EAR and RNI of mixed protein are 5% and 16% that are lower than the current proposed EAR and RNI {0.92 and 1.16 g/（kg.d）, respectively）, as determined by the nitrogen balance method. The respective EAR and RNI recommendations of 0.87 and 0.98 g/（kg.d） of mixed protein are estimated to be reasonable and suitable for Chinese young adults.

Genome-wide association studies reveal the genetic basis of amino acid content variation in tea plants

Tea is one of the most popular non-alcoholic beverages in the world,and free amino acids,especially theanine,make a major contribution to the umami taste of tea.However,the genetic basis of the variation in amino acid content in tea plants remains largely unknown.Here,we measured the free amino acid content in fresh leaves of 174 tea accessions over two years using a targeted metabolomics approach and obtained genotype data via RNA sequencing.Genome-wide association studies were conducted to investigate loci affecting the content of free amino acids.A total of 69 quantitative trait loci(–log10(P-value)>5)were identified.Functional annotation revealed that branched-chain amino acid aminotransferase,glutamine synthetase,nitrate transporter,and glutamate decarboxylase might be important for amino acid metabolism.Two significant loci,glutamine synthetase(Glu1,P=3.71×10^(−4);Arg1,P=4.61×10^(−5))and branched-chain amino acid aminotransferase(Val1,P=4.67×10^(−5);I_Leu1,P=3.56×10^(−6)),were identified,respectively.Based on the genotyping result,two alleles of CsGS(CsGS-L and CsGS-H)and CsBCAT(CsBCAT-L and CsBCAT-H)were selected to perform function verification.Overexpression of CsGS-L and CsGS-H enhanced the contents of glutamate and arginine in transgenic plants,and overexpression of CsBCAT-L and CsBCAT-H promoted the accumulation of valine,isoleucine and leucine.Enzyme activity assay uncovered that SNP1054 is important for CsGS catalyzing glutamate into glutamine.Furthermore,CsGS-L and CsGS-H differentially regulated the accumulation of glutamine,and CsBCAT-L and CsBCAT-H differentially regulated the accumulation of branched-chain amino acids.In summary,the findings in our study would provide new insights into the genetic basis of amino acids contents variation in tea plants and facilitate the identification of elite genes to enhance amino acids content.

Determination of basal ileal endogenous losses and standardized ileal digestibility of amino acids in barley fed to growing pigs

Background： Basal ileal endogenous amino acid（AA） losses（IAAend） and standardized ileal digestibility（SID） values of cereal grains, such as barley, are apparently underestimated when determined according to the nitrogen（N）-free method. Regression analysis between the dietary apparent ileal digestible content（c AID） and total crude protein（CP） and AA can be considered as alternative approach to obtain more accurate values for IAAendand SID of AA in cereal grains.Methods： Eight hulled barley genotypes were used, with barley being the only source of CP and AA in the assay diets. The diets contained 95 % as-fed of these eight barley genotypes each, ranging in CP content between 109.1 and 123.8 g/kg dry matter（DM）. Nine ileally T-cannulated barrows, average body weight（BW） 30 ± 2 kg, were allotted to a row-column design comprising eight periods with 6 d each and nine pigs. On d 5 and the night of d 6 of every period, ileal digesta were collected for a total of 12 h. The IAAend and the SID were determined by linear regression analysis between c AID and total dietary CP and AA.Results： There exist linear relationships between cA ID and total CP and AA（P 〈 0.001）. The IAAend of CP, Lys, Met, Thr and Trp amounted to 35.34, 1.08, 0.25, 1.02 and 0.38 g/kg DM intake（DMI）, respectively, which are greater compared to average IAAend determined previously under N-free feeding conditions. The SID of CP, Lys, Met, Thr and Trp was 90,79, 85, 79 and 86 %, respectively, and was greater when compared to tabulated values. Moreover, these SID values were greater than those reported in literature, based on correction of apparent ileal digestibility（AID） of CP and AA for their IAAendvalues. Summarized, the results of the present regression analysis indicate greater IAAendin barley-based diets compared to those obtained by N-free feeding.Conclusions： For low-protein feed ingredients like barley the regression method may be preferred over correction of AID values for their IAAenddetermined under N-free feeding conditions, as intercepts and slopes of the linear regression equations between cA ID and total dietary CP and AA provide direct estimates of IAAendand SID of CP and AA in the presence of the assay feed ingredient.

Involvement of sulfur assimilation in the low β subunit content of soybean seed storage protein revealed by comparative transcriptome analysis

The β subunit of soybean [Glycine max (L.) Merr.] seed storage protein is of great significance in sulfur-containing amino acid balance and soybean processing properties. The objective of this study was to elucidate the relationship between the β subunit and sulfur-containing amino acid composition, and the potential regulatory mechanism. The β subunit was independently accumulated in comparison with other major subunits (α/α′, acidic, basic, and A3) during seed filling, and a low level of β subunit content (BSC) was formed during the accumulation process. In low-BSC mature seeds, crude protein, oil content, and fatty acid composition were not changed, but sulfur-containing amino acids (Cys + Met) in the low- BSC seeds increased significantly (by 31.5%), suggesting that an internal regulatory mechanism within seed might be responsible for the rebalance of seed protein composition and that sulfur assimilation might be deeply involved in β subunit accumulation. Transcriptomic analysis revealed that genes involved in anabolism of cysteine, methionine, and glutathione were up-regulated but those involved in the catabolism of these compounds were down-regulated, suggesting a relationship between the elevation of methionine and glutathione and low BSC. Our study sheds light on seed composition in low BSC lines and on the potential molecular regulatory mechanism of β subunit accumulation, broadening our understanding of soybean seed protein synthesis and its regulation.

Determination of Amino Acids in an Individual Erythrocyteby Capillary Electrophoresis with Intracellular FITC-derivatization and Laser-induced Fluorescence Detection

A novel approach for analysis of amino acids in individual erythrocytes was established. In this method, the derivatization reagent was introduced into the living cells by electroporation. After derivatization, the amino acids in a single cell were determined by capillary electrophoresis with laser-induced fluorescence detection.

Proteomic analysis of energy metabolism and signal transduction in irradiated melanoma cells

AIM: To analyze proteomic and signal transduction alterations in irradiated melanoma cells. METHODS: We combined stable isotope labeling with amino acids in cell culture (SILAC) with highly sensitive shotgun tandem mass spectrometry (MS) to create an efficient approach for protein quantification. Protein protein interaction was used to analyze relationships among proteins. RESULTS: Energy metabolism protein levels were significantly different in glycolysis and not significantly different in oxidative phosphorylation after irradiation. Conversely, tumor suppressor proteins related to cell growth and development were downregulated, and those related to cell death and cell cycle were upregulated in irradiated cells. CONCLUSION: Our results indicate that irradiation induces differential expression of the 29 identified proteins closely related to cell survival, cell cycle arrest, and growth inhibition. The data may provide new insights into the pathogenesis of uveal melanoma and guide appropriate radiotherapy.

Effect of Adjunct Culture <i>Lactobacillus helveticus</i>(B02) on the Composition, Proteolysis, Free Amino Acids Release and Sensory Characteristics of Prato Cheese

At semi-industrial scale it was obtained two cheeses, one with aromatic culture (CHN) and the other with also L. helveticus (CHN + LH). Proteolysis during ripening was evaluated using proteolytic indices, electrophoretic profile Urea-PAGE and release of free amino acids. Sensory evaluation was performed by acceptability test. It was observed a 5-fold increase in the fraction corresponding to αs1-I-casein in the CHN cheese and 6.5 times in the CHN + LH cheese and 7.2 and 8.2 increase respectively, for total amino acids. Prato cheese with the adjunct culture showed higher acceptability in terms of overall aroma and flavor.

Analysis and Evaluation of Nutrient Composition in Different Tissues of the Wild Spot-fin Porcupine Fish(Diodon hystrix)

Nutrient compositions including proximate, fatty acids, amino acids, minerals and vitamins in tissues of the wild spot-fin porcupinefish(Diodon hystrix) were analyzed in detail to assess their nutrient value and to further explore the dietary requirements of this species farmed. Diodon hystrix individuals with similar body weight(284.6-358.5 g) and body length(15.9-21.8 cm) were collected from the South China Sea from Aug., 2015 to Aug., 2016. In general, skin and muscle possessed negligible fat contents(0.3-0.4 g/100 g wet weight) and low energy value(317.4-497.3 k J/100 g), while proteins(17.8-25.4 g/100 g wet weight) were found to be the richest component. The dominant amino acids in both muscle and skin of D. hystrix included histidine, cysteine, tyrosine, isoleucine, leucine and lysine, while the major mineral elements were calcium, magnesium, potassium,sodium, and phosphorus. It was apparent that D. hystrix had high content of amino acids, and the rate of essentia amino acids was accorded with FAO/WHO standard. This study illustrated that the wild D. hystrix would be a valuable natural source of protein, with high commercial potential, and especially, skin and muscle were more valuable. Moreover, the basic data from the present study would supply the fundamental reference for formulated diet and culture of this species.

Differentiation of bee pollen samples according to the apiary of origin and harvesting period based on their amino acid content

Bee pollen is currently one of the most widely consumed dietary supplements due to its high nutritional value and its potentially beneficial effects on health. Unfortunately, in recent years an increase in the fraudulent marketing of this product has been detected, mainly in terms of adulteration with pollen from other sources. This has made it necessary to seek new tools to ensure its authentication. Therefore, this study investigates the use of free amino acids as markers of the geographical origin and harvesting period of bee pollen. To demonstrate their potential as biomarkers, 72 samples from four apiaries (Pistacho, Tío Natalio, Monte and Fuentelahiguera), located in the same geographical area (Marchamalo, Guadalajara, Spain), were analyzed by liquid chromatography-fluorescence detection, with the data obtained undergoing canonical discriminant analysis. Variable amounts and numbers of free amino acids were found in the samples analyzed;proline predominated in all of them, in a concentration range of 298–569989 mg/kg. The differences observed in amino acid composition could be attributed to the flowering plants from which the bee pollen samples originated. In addition, it was possible to statistically assign over 75% of the samples to the corresponding apiary of origin, the best results being obtained for the Fuentelahiguera and Tío Natalio apiaries (100%);this classification was even superior in the case of the harvesting periods, as more than 90% of the samples were correctly assigned, and in one period (June) a 100% rate was obtained.

Identification of Geographical Origins for Blackcurrant Cultivars Based on Amino Acid Compositions

In this study,a total of 36 blackcurrant(Ribes nigrum L.)cultivars grown in the Northeast of China were selected,including 12 cultivars introduced from Russia,10 from Poland and the rest from local areas.The physicochemical properties and amino acid compositions of these varieties were studied,and the geographical origins of blackcurrants were tracked by multivariate statistical analysis.A total of 23 amino acids were detected in all cultivars,which were rich in glutamine,glutamate,aspartate,asparagine,α-alanine,γ-aminobutyric acid,valine and serine.The content of the total amino acids in these cultivars was from 31.21 mg•100 g-1 to 319.40 mg•100 g-1.Stepwise linear discriminant analysis(SLDA)was introduced to perform satisfactory categorization for blackcurrant cultivars,which achieved a success rate of 88.9%for the identification of geographical origins.These results suggested that the compositions of amino acids in blackcurrants could effectively predict geographical origins.

NMR-based metabolomics pro!le analysis in understanding metabolic variations of Bra LTP1-overexpressing Brassica napus

BraLTP1 is an important member of lipid transfer protein family in Brassica napus.The aim of current study was to detect, classify and follow variations in metabolite profiling of B. napus with overexpression of BraLTP1. In this study, metabolic change in leaves of BraLTP1-overexpressing B. napus plants (BraLTP1-22) and negative control (BraLTP1-22N) was investigated using nuclear magnetic resonance (NMR). Statistical strategy of principle component analysis (PCA) was performed to identify related difference metabolites. PCA score plots indicated not only high reproducibility of various treatments, but also significant difference of metabolite levels between different treatments. PCA loading plots indicated main responsible metabolites and a total of 50 metabolites were quantitatively determined. A wide range of metabolites were detected due to BraLTP1 overexpression,including biosynthesis and metabolism of sugars, amino acids, ammoniums compounds and organic acids. Furthermore, concentrations of 17 amino acids were determined for other set of samples by amino acids component analysis assay. The link between metabolite variations and phenotype were also discussed in BraLTP1-overexpressing B. napus.This work will help to gain insight into BraLTP1 function in B. napus in metabolism.