Differences in free and protein-bound tyrosine among potato genotypes and the relationship to internal blackspot resistance

Twelve potato clones were selected to represent the full range of internal blackspot response in order to determine the relationships between tuber protein, free tyrosine, and blackspot susceptibility. The blackspot reaction for each clone was consistent over five growing seasons, including tubers grown over a normal season (mature), and short season (immature) during one year. The blackspot index, determined by either an abrasive peel test or an impact bruise test, was highly correlated with the tyrosine content of the tubers (r = 0.90 p = 0.001 for the means of each clone over five location-years). Tubers with free tyrosine levels below 4 μmole/g dry weight consistently showed a resistant blackspot response. The relationship between tyrosine and blackspot susceptibility was also found in stolon and bud ends from five of the clones which represented the extremes of blackspot reaction and genetic diversity. Bud end samples of each of the clones had lower tyrosine content and a corresponding reduction in blackspot compared with stolon ends. Phenols, other than tyrosine, showed no consistent relationship to the blackspot reaction. There was a very high negative correlation between free tyrosine and estimated protein-bound tyrosine. R values ranged from ?0.85 to ?0.97 (p = 0.001) for mature tubers of the 12 clones over 4 growing seasons. Total tyrosine (free, plus protein-bound) remained relatively constant. There were no significant differences in mean total tyrosine content among the 12 clones over five location-years of testing; and there were no significant differences among the five growing seasons except for the short season (immature) tubers which were 14% lower in total tyrosine content. These results indicate a remarkably constant level of total tyrosine production in the twelve clones studied, that represented diverse genetic backgrounds. Mature tubers of all genotypes contained 26 ± 1 μmole/g dry weight total tyrosine. Partitioning of tyrosine between tuber protein and the free amino acid pool varied with genotype and appeared to be a major determinate of blackspot resistance.     

Characterisation and functional properties of Australian rice protein isolates

Albumin, globulin, glutelin and prolamin fractions were isolated from an Australian rice variety (cv. Langi) and characterised by yield, protein content and molecular weight profile using both capillary electrophoresis (SDS-CE) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The influence of pre-extraction enzymatic hydrolysis of starch and heating to 70 °C was also investigated, as was the extraction of the glutelin fraction without prior removal of the albumin and globulin fractions. Pre-extraction treatment affected mainly the albumin fraction, increasing dry matter yield but reducing protein content. SDS-CE was able to separate the protein fractions over a wider molecular weight range than SDS-PAGE, and the peaks from SDS-CE showed slightly higher molecular weight compared to equivalent bands from SDS-PAGE. The glutelin fraction extracted without prior removal of albumin and globulin fractions had different characteristics compared to those obtained by conventional extraction methods. Pre-extraction hydrolysis of starch did not significantly affect the emulsifying, foaming and gelling properties of extracted protein. Although rice glutelin had poor solubility, emulsifying and foaming properties in aqueous systems, it had good gelling properties which could be important for food applications.     

Amino acid compositions of different protein fractions in developing grains of NP 113 barley and its high lysine Notch-2 mutant

The percent distributions of protein fractions namely albumin + globulin, prolamine and glutelin were studied in developing grains of NP 113 barley and its high lysine mutant Notch-2. During development the percentage of albumin + globulin fraction decreased in NP 113, while those of prolamine and glutelin remained unchanged. The increase in prolamine was substantial from 24 to 31DAA. In Notch-2 the trend followed by albumin + globulin and prolamine was like that in NP 113, while the glutelin fraction showed an increase as compared to 10 DAA. The percent of albumin + globulin was slightly higher in Notch-2 as compared to NP 113. The absolute amount (mg/grain) of all the protein fractions increased during development in both NP 113 and its mutant Notch-2. During the grain development the prolamine content was substantially lower in the mutant than in the parent NP 113. The albumin + globulin content per endosperm was in general also higher in NP 113 than Notch-2. Amino acid analysis of the protein fractions did not reveal significant changes in lysine between NP 113 and Notch-2. Thus, the improvement in lysine in the mutant is primarily due to reduced synthesis of the prolamine fraction and not due to an increase in lysine in the mutant hordein fraction. Part of the improvemenht in lysine may also be due to increase in the percentage of albumin + globulin fractions which is lysine rich.Part of the Ph.D thesis submitted by S. Joshi to P.G. School, IARI, New Delhi.     

Changes in nitrogenous and other chemical constituents,protein fractions and in vitro protein digestibility of germinating fluted pumpkin ( Telfairia occidentalis Hook) seed

The effect of 7 days of germination on levels of nitrogenous and other nutrition related parameters, protein fractions and in vitro protein digestibility of fluted pumpkin (Telfairia occidentalis) seed was studied. The non-protein nitrogen gradually increased and the protein nitrogen content decreased during germination. Albumin and globulin fractions were found to be the major seed proteins of fluted pumpkin seeds, constituting about 58.6% of the total protein of the ungerminated (raw) seeds. The protein fractions, albumin and glutelin, were observed to increase by 61.5% and 57.0%, respectively, while a 54.6% decrease was noted in the prolamine fraction. The globulin fraction increased at the beginning of germination but decreased at the end. Germination significantly (p 0.05) increased the crude protein, nitrogen solubility and in vitro protein digestibility but decreased the fat, phytic acid and polyphenol contents of the seeds.     

椰麸组分蛋白的氨基酸组成、结构与乳化性分析

为提高椰子加工副产物的利用,本研究从椰麸中依次提取4种组分蛋白,包括清蛋白、球蛋白、谷蛋白-1和谷蛋白-2.分析这4种组分蛋白的氨基酸组成,并利用SDS-PAGE技术分析其亚基组成和分子量.同时分析其表面疏水性,并研究不同离子强度对椰麸各组分蛋白的乳化性和乳化稳定性的影响.结果表明,椰麸蛋白质中清单白、球蛋白、谷蛋白-...     

Nutritional potential ofVigna minima (Roxb.) Ohwi and Ohashi:

Quantitative variation in different fractions of seed proteins and their amino acid levels in populations ofVigna minima (Roxb.) Ohwi and Ohashi and inV. umbellata cv. IC 1568 — the rice bean — were investigated. Globulin I fraction, together with globulin II, constitutes 38 to 54 per cent of the total seed protein. The alkali soluble (glutelin) fraction is the second largest fraction. Both these fractions show broad range of variation, suggesting a broad genetic base. The profiles are population specific; the coastal population, which contains higher seed protein also possesses maximum levels of globulin I and glutelin fraction suggesting its potentiality for breeding lines with high protein content, high nutritive value, and salt tolerance. Protein content is positively correlated with globulin I and glutelin fractions, which are in turn positively correlated with each other. The amino acid profiles are specific not only to the fractions but also to the populations. The range of variation in the levels of all amino acids in different fractions is broad suggesting substantial genetic diversity. The average levels of lysine and sulphur amino acids are high in globulin I and glutelin fractions.     

Transglutaminase polymerisation of buckwheat (Fagopyrum esculentum Moench) proteins

Recently, screening of transglutaminase (TGase) treatment on several gluten-free cereals revealed significant improvements on the baking performances of buckwheat flour by promoting protein networks. In this study, the impact of TGase on the protein fractions of buckwheat flour was investigated in order to better understand the activity and specificity of the enzyme. Albumin, globulin, prolamin and glutelin fractions were extracted from the flour and incubated with TGase. Capillary electrophoresis, two dimensional (2D) gel electrophoresis and size exclusion chromatography (SEC) were performed on each fraction. Capillary electrophoresis and 2D gel electrophoresis revealed that buckwheat main storage proteins, i.e. 2S albumin, 13S and 8S globulin, were cross-linked after TGase treatment. SEC showed the presence of high molecular weight (HMW) protein polymers in the TGase-treated albumin and globulin fraction. Analysis of the amino acid composition of the fractions revealed high amounts of glutamine and lysine residues in all fractions. In conclusion, the increase in the average molecular weight of buckwheat proteins and the formation of HMW protein polymers after TGase treatment are responsible for the improved functionality of buckwheat flour in terms of breadmaking potential. The enzyme was revealed to be not fraction specific as all fractions were TGase-reactive.     

Transglutaminase polymerisation of buckwheat (Fagopyrum esculentum Moench) proteins

Recently, screening of transglutaminase (TGase) treatment on several gluten-free cereals revealed significant improvements on the baking performances of buckwheat flour by promoting protein networks. In this study, the impact of TGase on the protein fractions of buckwheat flour was investigated in order to better understand the activity and specificity of the enzyme. Albumin, globulin, prolamin and glutelin fractions were extracted from the flour and incubated with TGase. Capillary electrophoresis, two dimensional (2D) gel electrophoresis and size exclusion chromatography (SEC) were performed on each fraction. Capillary electrophoresis and 2D gel electrophoresis revealed that buckwheat main storage proteins, i.e. 2S albumin, 13S and 8S globulin, were cross-linked after TGase treatment. SEC showed the presence of high molecular weight (HMW) protein polymers in the TGase-treated albumin and globulin fraction. Analysis of the amino acid composition of the fractions revealed high amounts of glutamine and lysine residues in all fractions. In conclusion, the increase in the average molecular weight of buckwheat proteins and the formation of HMW protein polymers after TGase treatment are responsible for the improved functionality of buckwheat flour in terms of breadmaking potential. The enzyme was revealed to be not fraction specific as all fractions were TGase-reactive.     

The effect of heat treatment on the soluble protein content of oats

The effect of heat treatment on the soluble protein content in oat groats (Kerstin commercial variety) was evaluated using asymmetric flow field-flow fractionation (AF4) in combination with online multiangle light scattering (MALS) and UV detection. The AF4 method was used to separate the monomeric proteins from globulin hexamer and aggregate proteins and β-glucan polysaccharides in the soluble oat protein fraction. The total amount of soluble protein (with respect to total protein) was reduced to 35.7 ± 4.5 wt. % in heat treated oats from 74.6 ± 5.3 wt. % in non-heat treated oats. The ratio of monomeric to globulin hexamer and aggregate proteins was reduced from 1.82 to 1.48 as a result of heat treatment. Sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed the selective elimination of protein bands associated with the albumin and prolamin protein fractions as a result of heat treatment. These results were supported through amino acid analysis by cation exchange chromatography coupled with UV detection which revealed a reduction in amino acid residues associated with prolamin. The globulin proteins were found to be less sensitive to heat treatment.     

Fundamental study on protein changes taking place during malting of oats

During the malting process, storage proteins are degraded by proteolytic enzymes into small peptides and amino acids. The activity of these enzymes was measured during malting of oats and was found to be increased. To quantify proteolytic degradation, proteins of unmalted, germinating and malted grains were fractionated. After extracting the oat proteins (Osborne fractionation), protein fractions were analysed using a Lab-on-a-Chip technique, which separates the proteins – based on their molecular weight – by capillary electrophoresis. This new technique for the analysis of proteins was supported by using two-dimensional gel electrophoresis. In addition, amino acid analysis was carried out. In general a degradation of proteins to small peptides and amino acids could be observed in the globulin, prolamin and glutelin fractions. In the albumin fraction a protein increase was observed, which is due to the fact that this fraction contains the majority of the metabolically active proteins. Amino acid analysis supported the observation of increased protein amount in the albumin fraction and decreased protein amounts in the other fractions. Some proteins, which have not been described in the literature, were detected in the albumin and glutelin fraction, since Lab-on-a-Chip technique allows detection of proteins with low molecular weights of 4.5 kDa.     

Antioxidative properties of partially purified barley hordein, rice bran protein fractions and their hydrolysates

Antioxidative properties of proteins from barley and rice bran and their hydrolysates were examined. Three major hordein fractions of barley, B, C and D hordeins, were partially purified by gel filtration. Albumin, globulin, prolamin and glutelin fractions of rice bran were fractioned by the Osborne method. Hydrolysates of these protein fractions were prepared by digesting with pepsin followed by trypsin. Antioxidant properties in terms of antioxidative activity against linoleic acid peroxidation and reducing activity without the lipid adjuvant were investigated. The globulin fraction from rice bran protein revealed the strongest antioxidative activity throughout the incubation time of 7 days (p ≤ 0.05). The albumin fraction of rice bran protein showed the highest reducing activity (6964 μmol of Fe²⁺) followed by globulin, prolamin, glutelin and hordein fractions with activities of 2904, 2017, 1809 and 1333 μmol of Fe²⁺, respectively (p ≤ 0.05). Partially purified C hordein exhibited the highest reducing activity compared with B and D hordeins. Protein hydrolysates obtained after digestion with pepsin and trypsin exhibited much greater antioxidative, as well as reducing, activities than those from before digestion.     

Effect of fermentation on sorghum protein fractions and in vitro protein digestibility

Changes in pH, titratable acidity, total soluble solids and proteins ofDabar sorghum (Sorghum bicolor (Linn) Moench.) during naturalfermentation at 37 ^°C for up to 36 h were monitored. The pH ofthe fermenting material decreased sharply with a concomitant increase in the titratable acidity. Total soluble solids increased with progressivefermentation time. The crude protein and non-protein nitrogen slightlyincreased during the last stages of fermentation. The in vitroprotein digestibility markedly increased as a result of fermentation.The globulin plus albumin fractions increased significantly (p 0.05)during the first 8 h of fermentation. Kaffirin fraction decreasedduring the first 8 h of fermentation but increased sharply as fermentationprogressed. Cross-linked kaffirins fluctuated during the fermentationprocess. Glutelin like protein, which was the minor fraction, trueglutelins, the second most abundant fraction, together with non-extractableproteins fluctuated during the fermentation process.     

Distribution of seed protein fractions and amino acids in different anatomical parts of chickpea (Cicer arietinum L.) and pigeonpea (Cajanus cajan L.)

Studies on protein fractionation in seed coat, embryo, cotyledons and whole seed were made to observe the differences, if any, between chickpea and pigeonpea. Results indicated that globulin was the major fraction of embryo and cotyledons of these legumes. Seed-coat nitrogen was observed to be mostly comprised of nonprotein nitrogen and glutelin fractions and thus differed from other components in both chickpea and pigeonpea. The albumin fraction of cotyledons of both crops had the highest concentration of sulphur amino acids, methionine and cystine. Glutelin contained a considerably higher concentration of methionine and cystine than did globulin in chickpea and pigeonpea. This suggests that lines with higher glutelin should be identified to improve their protein quality. The amino acid compositions of different seed components did not show large differences between these two pulse crops.Submitted as JA no. 180 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT).     

Nutritional assessment of seed proteins in rice bean [Vigna Umbellata (Thumb.) Ohwi and Ohashi

Seeds of three rice bean accessions had 17.26 to 21.42% protein, 3.46 to 4.03% fat, 61.09 to 64.73% carbohydrates 3.99 to 4.58% ash and 5.22 to 7.43% fiber (dry weight basis).The most limiting amino acids in the seed meal, albumin and globulin fractions, were methionine and cysteine with chemical scores of these fractions being 38% to 59%. The amino acid pattern of globulin and seed meal were similar.Thein vitro protein digestibility (IVPD) ranged from 82 to 86% for the seed meal, 86 to 88.5% for the albumin and 75.9 to 83.3% for the globulin. Relative nutritive values (RNV) of raw mature seed of two accessions were 22.6% and 42.4% and increased to 55.6% to 79.4% after boiling and roasting.Part of MS thesis of the senior author.     

醋制对黑豆蛋白含量及多肽ACE抑制活性的影响

为考察醋制对黑豆蛋白含量及多肽ACE抑制活性的影响,为对黑豆醋浸过程中蛋白组分及含量变化进行分析,并对相应ACE抑制肽活性进行测定。黑豆经醋浸不同时间后,采用顺序抽提法提取清蛋白、球蛋白、醇溶蛋白和谷蛋白,并采用凯氏定氮法及SDS-PAGE电泳进行定量和定性分析;各蛋白组分经胃蛋白酶水解,经超滤(截留分子量3 k D)后收集滤液,获得多肽组分,RP-HPLC法进行ACE抑制活性评价。结果表明:经醋浸14 d后,黑豆总蛋白含量变化幅度小于±0.5%;清蛋白含量由55.13%(0 d)降低至9.61%(14 d);球蛋白由7.04%(0 d)增加到20.34%(14 d);醇溶蛋白由0.81%(0 d)增加到1.72(14 d);谷蛋白由21.11%(0 d)增加到59.45%(14 d),含量最高。醋浸处理降低了清蛋白源多肽的ACE抑制活性,但提高了球蛋白、醇溶蛋白及谷蛋白多肽的ACE抑制活性,其中,谷蛋白多肽抑制率由18.18%(0 d)增加至37.58%(14 d),抑制活性升高。醋浸可改变黑豆各蛋白组分的含量和对应多肽的ACE抑制活性,其中,谷蛋白含量及其多肽ACE抑制活性均有增加。研究结果表明可进一步采用分离纯化技术从谷蛋白多肽中获得高活性降压肽。     

Seed protein fractions and amino acid composition in gram (Cicer arietinum)

Six chickpea strains were analysed for their protein content and various protein fractions. The protein content ranged from 20.9–25.27%. Albumin, globulin, prolamin and glutelin contents ranged from 8.39–12.31%; 53.44–60.29%; 3.12–6.89% and 19.38–24.40% respectively. Salt soluble proteins (albumin + globulin) and globulins resolved into 19–23 bands whereas albumin proteins resolved into 30–34 bands. The molecular weights of various polypeptides ranged from 10–91 kD. Amino acid analysis of total proteins revealed that glutamic acid was present in maximum concentration followed by aspartic acid and arginine. Just like other pulse proteins, chick pea proteins were also found deficient in sulphur containing amino acids.     

我国江浙地区粳稻品种间的植酸与蛋白质组分差异及其相关性

选用我国江浙地区近期育成的晚粳稻品种（系）29个,对它们在浙江杭州相同栽培条件下籽粒中的植酸、蛋白质含量与其组分差异及其相关性进行了分析。供试29个品种(系)植酸含量的平均值为0.868%,变化幅度为0.699%~1034%,其中秀水系列品种的植酸含量一般较低,而武育粳系列和淮字系列品种（系）的植酸含量则相对较高;供试29个品种材料总蛋白质含量的平均值为8.722%,其含量大致呈正态分布。在4种蛋白质组分中,谷蛋白、球蛋白和清蛋白在品种间的变异系数较大,而醇溶蛋白则相对较小,且在分布频率上也表现出品种类型间的差异特征;除谷蛋白与总蛋白质之间存在着极显著的正相关外,供试29个品种(系)的植酸含量与蛋白质含量及4种蛋白质组分含量间并无密切的关系。     

Effect of cooking on cowpea protein fractions

Nine cowpea cultivars obtained from Wad Medani Research Station were used in this study. The variation in protein fraction was: albumins 4.0–12.0, globulins 65.6–79.7, prolamins 1.4–2.2, G₁-glutelins 0.9–3.0, G₂-glutelins 1.4–2.9. G₃-glutenins 9.1–14.0 and insoluble protein 0.5–3.0%. Two cowpea cultivars, H8-14 and CB-46, selected for their high albumin content, were cooked in 150 ml of boiling distilled water under reflux for 45 minutes. The protein fractions in the cooked seeds were determined. Results indicated that albumin and globulin fractions decreased significantly (p0.05) for both cultivars after cooking. This decrease was accompanied by a significant increase in G₃-glutelin fractions.     

Influence of moisture stress during growth on14CO2 fixation and translocation inSolanum tuberosum L.

Moisture stressing of potato plants resulted in reduced¹⁴CO₂ fixation and translocation of labelled photosynthates from leaves to tubers. A majority of the label was recovered in the sugar fraction of both leaves and tubers. The amount of labelled¹⁴C recovered in the organic acid fraction of tubers of normally irrigated plants was significantly higher than in tubers which had been moisture stressed. In the other fractions, the differences were not significantly different. Injection of uniformly labelled sucrose into the basal portion of attached tubers of stressed and non-stressed plants showed greater translocation of labelled carbon by tubers of stressed plants from basal to the apical portion and also into the stems as compared to non-stressed plants.     

Biochemical characteristics of potato clones differing in blackspot susceptibility

The relationships between enzymatic discoloration and selected biochemical characteristics of tubers from nine clones were determined in an attempt to explain varietal differences in blackspot susceptibility. These nine clones represent a wide range of blackspot susceptibility as determined by two discoloration indices after abrasive peeling. Enzymatic discoloration for combined data from the 1983 and 1984 growing seasons was highly correlated with total phenol (r=0.89) and tyrosine (r=0.85) concentrations. Conversely, discoloration was negatively correlated with soluble protein in 1983 (r=-0.78) and 1984 (r=-0.77). Polyphenoloxidase (PPO) activities, apparent Michaelis constants and maximum velocity values, from crude and partially purified enzyme preparations, were not significantly correlated with enzymatic discoloration. Varietal susceptibility to enzymatic discoloration appears to be related to concentrations of endogenous phenolic substrates of PPO, rather than to differences in the quantity or properties of the PPO enzyme.