玉米低植酸突变体的营养品质分析

对比研究了8个玉米低植酸(Low phytic acid,lpa)突变体及其相应野生型(Wild Type,WT)的主要营养品质及其营养组分,结果显示:与野生型相比,各突变体总磷含量基本不变,无机磷含量均成倍增加,总淀粉含量显著下降,直链淀粉和粗脂肪含量变化不显著,粗蛋白含量在部分突变体中显著上升;多数氨基酸含量在突变体中有所提高,赖氨酸含量除Q319-lpa和X178-lpa下降外均不同程度上升;铁、锰、铜、锌、硒等微量元素含量变化不显著。研究表明lpa突变对玉米营养品质的改良和生物学效价的提高具有有益的作用。     

Electrophoretic method for the identification of a haze-active protein in grape seeds

A simple, fast, and more selective approach is presented in this study for the identification of haze-active proteins. Grape seed proteins were unfolded by 1% SDS and then interacted with different amounts of tannin at 4 degrees C, followed by gel electrophoresis. It was found that the intensity of the band at 45 kDa was decreased as tannins increased. The amino acid composition of this isolated 45-kDa protein was higher in proline (9.49%) than the average proline content of total grape seed proteins (4.85%). To verify the selectivity of the proposed method, a globular protein (bovine serum albumin, BSA) and a proline-rich protein (gelatin) were selected and used in the model system. As expected, gelatin was removed as it reacted with the increasing added tannins, whereas BSA did not. These results showed that it is possible to identify haze-active proteins by modulating the accessibility of protein to tannins, suggesting this new method can be used by the beverage industry to trouble-shoot haze problems and for quality control.     

Assessment of the protein quality of 15 new northern adapted cultivars of quality protein maize using amino acid analysis

Amino acid determinations were carried out on 15 new northern adapted cultivars of quality protein maize (QPM) containing opaque-2 modifier genes to ascertain whether their amino acid scoring patterns could be used to select high-lysine QPM genotypes and to assess their protein quality. Total protein in these cultivars ranged from 8.0 to 10.2% compared to two commercial maize varieties, Dekalb DK435 (7.9%) and Pioneer 3925 (10.3%). Four of these QPM genotypes, QPM-C26, QPM-C21, QPM-C79, and QPM-C59, contained high levels of lysine (4.43-4.58 g of lysine/100 g of protein), whereas the remaining varied from 3.43 to 4.21 g of lysine/100 g of protein, compared to Dekalb DK435 and Pioneer 3925, which contained 2.9 and 3. 1 g of lysine/100 g of protein, respectively. Although lysine is the first limiting amino acid in QPM inbreds, the high-lysine QPM genotypes may supply approximately 70.2-72.6% of human protein requirements, compared to 46.2% for Dekalb DK435 and 50.1% for Pioneer 3925, 55-63% for oats, and 59-60.3% for barley. Northern adapted QPM genotypes may have the potential to increase their lysine content even further, either by an increase in specific high-lysine-containing nonzein proteins, such as the synthesis of factor EF-1a, or by a further reduction in the 19 and 22 kDa alpha-zein in the endosperm or both. This knowledge could assist maize breeders in the selection of new high-performance QPM genotypes with improved protein quality and quantity.     

Improved organoleptic and nutritive properties of bakery products supplemented with amino acid overproducing Saccharomyces cerevisiae yeasts

Spontaneous yeast mutants isolated in continuous culture as resistant to toxic amino acid analogues, able to increase up to 40 times their free amino acid pool of Thr, up to 160 times their pool of Met, or up to 20 times their pool of Lys, were characterized with regard to properties of industrial interest. Growth rate, mu (h(-1)), and biomass yield, Y (g/L), of the amino acid overproducing mutants (AA(S)) were in many cases similar to those of the wild type, whereas their free amino acid content was substantially increased in laboratory and industrial media (molasses). Doughs fermented with 3% baker's yeast and 0.5% AA(S) mutants produced bakery products that displayed texture similar to those fermented with 3.5% baker's yeast, but the former had a considerable improvement of their taste and aroma. On the other hand, bread content of the essential amino acids Lys, Met, and Thr provided by yeast was also increased.     

Hypochlorous and peracetic acid induced oxidation of dairy proteins

Hypochlorous and peracetic acids, both known disinfectants in the food industry, were compared for their oxidative capacity toward dairy proteins. Whey proteins and caseins were oxidized under well controlled conditions at pH 8 as a function of the sanitizing concentration. Different markers for protein oxidation were monitored. The results established that the protein carbonyl content was a rather unspecific marker for protein oxidation, which did not allow one to differentiate the oxidant used especially at the lower concentrations. Cysteine, tryptophan, and methionine were proven to be the most vulnerable amino acids for degradation upon hypochlorous and peracetic acid treatment, while tyrosine was only prone to degradation in the presence of hypochlorous acid. Hypochlorous acid induced oxidation gave rise to protein aggregation, while during peracetic acid induced oxidation, no high molecular weight aggregates were observed. Protein aggregation upon hypochlorous acid oxidation could primarily be linked to tryptophan and tyrosine degradation.     

大杯香菇辐射选育新株系子实体蛋白质构成的多元回归与聚类分析

以20个大杯香菇品种和辐射选育新株系为材料, 分析了大杯香菇品种和辐射选育新株系蛋白质及其构成的17种氨基酸含量间的多元相关、回归和聚类关系, 并采用生物统计方法分析品种和新株系的蛋白质及各种氨基酸间的关系。结果表明: 大杯香菇辐105号新株系子实体中天门冬氨酸、苏氨酸、甘氨酸、丙氨酸、胱氨酸、缬氨酸、异亮氨酸、苯丙氨酸、蛋白质含量最高, 与部分品种和新株系子实体间差异显著或极显著, 且大多数氨基酸间及其与蛋白质含量之间存在显著或极显著正相关关系; 各种氨基酸与蛋白质含量之间分别有5种呈一次曲线关系, 10种呈二次曲线关系, ２种呈三次曲线关系; 采用系统聚类的方法可将蛋白质及17种氨基酸含量聚为4个大类, 对每类中的典型性状如精氨酸、亮氨酸、胱氨酸和蛋氨酸含量进行优化, 可有效提高大杯香菇品质育种中蛋白质构成的改良效率。     

Nitrogen lowers the sulfur amino acid content of soybean (Glycine max [L.] Merr.) by regulating the accumulation of Bowman-Birk protease inhibitor

Soybeans in general contain 35-40% protein. Efforts are underway to increase further this protein content, thus enhancing their nutritive value. Even though higher protein is a desirable characteristic, whether such an increase will be accompanied by enhanced protein quality is not known. Soybean protein quality could be significantly improved by increasing the concentration of the sulfur-containing amino acids, cysteine and methionine. To ascertain if a correlation existed between protein quantity and quality, a comparison of the amino acids of soybeans differing in protein content was made. Soybeans with higher protein content had a significantly lower percentage of sulfur amino acids, while those with lower protein exhibited a higher content of cysteine and methionine. Nitrogen application elevated the protein content but lowered that of the sulfur amino acids. Transmission electron microscopy examination of thin sections of low protein soybean seeds revealed several protein storage vacuoles that were partially filled with storage proteins. Fluorescence two-dimensional difference gel electrophoresis of soybean seed proteins revealed that nitrogen application favored the accumulation of the beta-subunit of beta-conglycinin while decreasing the accumulation of Bowman-Birk protease inhibitor (BBI), a protein rich in cysteine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 60% 2-propanol-extracted proteins showed a drastic reduction in the accumulation of BBI with increasing protein content. Northern blot analysis indicated that nitrogen had a negative influence on the expression of the BBI gene. Our results indicate that the negative correlation between total protein and sulfur amino acid content is mostly mediated by the differential accumulation of BBI.     

Evaluation of amino acid analysis as reference method to quantitate highly purified proteins

The applicability of amino acid analysis for accurate quantitation of reference standard preparations of proteins has been evaluated. This approach is very useful since, in addition to absolute quantitative information, it also provides a measure of composition, partial identity, and purity in a single experiment. Comparisons with Kjeldahl nitrogen assay and/or UV measurements shows that amino acid analysis is reliable for the quantitation of small-to-medium size proteins in the molecular weight range of 6-22 kDa. For larger proteins such as immunoglobulins (150 kDa), amino acid analysis may "underestimate" the total protein concentration. These results also show the effect of recovery of individual residues on protein quantitation. As expected, the recovery of more than one stable residue could be used to calculate total protein content of samples, which is in good agreement with the results obtained by Kjeldahl nitrogen assay. However, the protein concentrations calculated from the total mass of the recovered residues appear to give relatively low estimates in almost all cases. Thus, it is concluded that amino acid analysis is an appropriate reference method only when stable residues are employed for quantitation, particularly for highly purified proteins of rDNA origin.     

蚕豆发芽过程中蛋白质和抗氧化能力的变化

为更好地开发利用蚕豆资源,研究蚕豆发芽过程中蛋白质及蛋白抗氧化能力的变化规律,采用凯氏定氮法测定发芽蚕豆的蛋白质含量,采用氨基酸分析仪测定氨基酸组分含量,运用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术观察发芽蚕豆蛋白组成的变化,通过测定发芽蚕豆蛋白的1,1-二苯基-2-三硝基苯肼(DPPH)自由基和2,2'-联氮双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)自由基的清除能力分析其抗氧化能力。结果表明,蚕豆发芽过程中蛋白和氨基酸含量升高,促进部分蚕豆蛋白由大分子量转化分解成小分子量,提高了蚕豆蛋白的抗氧化能力,在蚕豆发芽第9天时蛋白质含量达到34.1 g·100g^-1、氨基酸含量为29.16 g·100g^-1,均达到最大值,蚕豆蛋白的自由基清除能力达到最强。综上,蚕豆的发芽过程可以增加蚕豆的营养物质成分含量,增强蚕豆的抗氧化能力。本研究结果为蚕豆蛋白、蚕豆芽等功能食品的开发利用提供了参考依据。     

Influence of cultivar and fruit ripening on olive (Olea europaea) fruit protein content, composition, and antioxidant activity

Proteins of olive fruit mesocarp are not very well-known at present. However, they have been shown to pass, at least partially, to the olive oil during its elaboration and therefore might be contributing to some of the special characteristics of this vegetable oil. In this study, protein content and composition were determined in olive fruits, cv. Arbequina and Picual, at three stages of ripening: green, spotted, and purple. Mesocarp proteins constituted 1.3-1.8% of the dry weight of the olive fruit, and cultivar and fruit ripening did not produce important changes in mesocarp protein content or composition. In addition, this composition was also similar to the amino acid composition of a 4.6-kDa polypeptide, which is the major protein component of olive oils and of oil bodies of olive fruit mesocarp, suggesting that this polypeptide is likely to be a major component of mesocarp proteins. There was, also, a relationship between the oil content of the olive fruit and the protein content determined, suggesting a stabilizing function of these proteins in the oil bodies of the olive fruit, analogously to the role suggested for oleosins. This stabilizing function does not seem to be extended to olive oils because when the polypeptides isolated were added at 20 ppm to soybean oil, the stability of the oil increased only slightly, suggesting that if these compounds play some role in the stability of the oils, this should be mostly a consequence of the possible interactions among these protein components and other olive oil antioxidant constituents.     

葵花粕中分离蛋白的成分及特性

该文以水酶法提取葵花籽油后的葵花粕为原料,通过盐提酸沉法提取其中的分离蛋白,并对其进行组分分析、氨基酸成分分析、电泳图谱分析和凝胶色谱分析,同时对其溶解性、吸水性、吸油性、乳化性和起泡性等功能特性进行研究。结果显示葵花籽分离蛋白的氨基酸组成与FAO必须氨基酸需要量模式相比赖氨酸含量较低,功能特性好于或接近大豆分离蛋白。葵花分离蛋白含有3个主要组分,其分子量分别为380×10³,100×10³和27×10³,绿原酸与分离蛋白紧密结合。     

EMS诱变花生珍珠红1号创制优良新种质

为创制花生优良新种质,以花生品种珍珠红1号的干种子为材料,采用浸泡法开展甲基磺酸乙酯(EMS)诱变研究。结果表明,以半致死剂量为选择标准,适宜诱变条件为0.8% EMS处理10 h;筛选获得9个优良M₅突变系,其中4个表现出产量高、品质优、综合性状优良等特点。上述4个M₅突变系干荚果产量较对照(珍珠红1号)增加10.93% ~ 26.78%,出仁率均高于对照;在品质方面,诱珍珠红1号2Ⅱ/19油酸含量为56.68%、油酸/亚油酸比值(O/L)为1.92,分别较对照提高8.44个百分点和0.57;诱珍珠红1号2Ⅲ/8亚油酸含量为39.05%,较对照提高3.28个百分点;诱珍珠红1号2Ⅲ/10粗脂肪含量为54.52%,较对照提高1.50个百分点;诱珍珠红1号3Ⅰ/8油酸含量为53.70%、O/L为1.71,分别较对照提高5.46个百分点和0.36。ISSR分子标记结果表明,9个优良M₄突变系与珍珠红1号在分子水平上均存在明显差异,突变系之间在分子水平上也存在明显差异。本研究结果为花生遗传改良和功能基因研究提供了优良的新种质,为花生EMS诱变育种研究提供了参考。     

Environmentally induced changes in amino acid composition in the grain of durum wheat grown under different water and temperature regimes in a Mediterranean environment

Amino acid composition is an important feature in determining the nutritional value of wheat grain for human and animal diets. Environmental conditions are known to influence protein quantity as well as grain production and, in turn, amino acid composition. In this study, grain yield, protein content, and amino acid composition were determined in 10 durum wheat genotypes under three water and temperature regimes in a Mediterranean environment. The highest value for grain-protein content (15.7%) was found in the warmer and driest environment and the lowest (12.8%) in the irrigated environment. Although amino acid composition showed significant variation for all genotypes, with the exception of arginine and cysteine, major changes in amino acid composition were caused by environmental conditions and in particular by water availability and temperature during the grain-filling period, which significantly altered the duration of grain development. The amino acids with the highest percentage of variation between environments were tyrosine (26.4%), lysine (23.7%), methionine (20.3%), threonine (19.3%), and valine (15.6%), whereas phenylalanine (5.1%), glycine (6.4%), and aspartic acid (6.8%) showed the least variation between environments. Whereas the content of glutamine, phenylalanine, and proline increased with the decrease in grain-filling duration, the remaining amino acids tended to diminish, presumably because high temperature and drought favored the deposition of gliadins (proteins particularly rich in glutamine and proline), to the detriment of albumins and globulins (proteins especially rich in threonine, lysine, methionine, valine, and histidine). Despite the negative correlations found between the percentage of protein and its content in essential amino acids, the results indicate that reductions in lysine per unit of food were not very pronounced (0.32 to 0.29 g/100 g of flour) with increases of up 22.7% in grain-protein content, whereas threonine did not change and valine even slightly increased.     

Bioengineering of Leuconostoc mesenteroides glucansucrases that gives selected bond formation for glucan synthesis and/or acceptor-product synthesis

The variations in glucosidic linkage specificity observed in products of different glucansucrases appear to be based on relatively small differences in amino acid sequences in their sugar-binding acceptor subsites. Various amino acid mutations near active sites of DSRBCB4 dextransucrase from Leuconostoc mesenteroides B-1299CB4 were constructed. A triple amino acid mutation (S642N/E643N/V644S) immediately next to the catalytic D641 (putative transition state stabilizing residue) converted DSRBCB4 enzyme from the synthesis of mainly α-(1→6) dextran to the synthesis of α-(1→6) glucan containing branches of α-(1→3) and α-(1→4) glucosidic linkages. The subsequent introduction of mutation V532P/V535I, located next to the catalytic D530 (nucleophile), resulted in the synthesis of an α-glucan containing increased branched α-(1→4) glucosidic linkages (approximately 11%). The results indicate that mutagenesis can guide glucansucrase toward the synthesis of various oligosaccharides or novel polysaccharides with completely altered linkages without compromising high transglycosylation activity and efficiency.     

辐照大豆诱发突变及其突变体的检测

不同剂量的γ射线、电子束单因子及其与不同浓度的苯甲酰胺复因子处理大豆种子，均能诱发其生育期、孕性和株高等农世性状的突变，同时还能诱发蛋白质、氨基酸等品质突变，而且复因子处理较单因子处理的突变频率高。此外，筛选出的不同类型的突变体，其过氧化物酶海性及过气体物酶同工酶酶谱均有明显差异。因此，可应用过氧化物酶等同工酶酶谱分析法对突变体进行检测。     

Nutritional responses of rats to diets based on chickpea ( Cicer arietinum L.) seed meal or its protein fractions

The aim of this study was to isolate the protein fractions from chickpea, var. IAC-Marrocos, as well as to evaluate its in vivo nutritional protein quality. Among the proteins, albumins showed better nutritional value in the in vivo assays and amino acid contents, despite their higher trypsin inhibitor contents. Trypsin inhibitors were found to be heat labile in all samples, but the digestibility results for unheated and heated flour and albumins suggest that their contents are not very decisive. The PER values for casein (not supplemented) were very similar to those of heated flour and unheated or heated albumin and total globulins. The albumin and glutelin fractions showed the best results for PDCAAS, however, lower than those of casein. Despite the high digestibility of the globulin the very low essential amino acid content lowered its PDCAAS, and it had the lowest values.     

人GJB2分子进化与耳聋遗传效应的关联性分析

间隙连接蛋白β2（GJB2）基因突变与遗传性非综合征性耳聋密切相关,其广泛的突变类型及特异性的热点突变被认为是一种独特的致聋基因。本研究应用生物信息学方法对17个物种的GJB2蛋白进行了系统发育、保守性、跨膜区结构、三维结构和错义突变的分析,并结合已有报道的实验结果进行关联性分析。分析预测获得了166个固定的氨基酸位点、2个非保守区以及2个空间结构保守位点;关联性分析证实发生在保守位点的突变致病性高,非保守区突变的概率致病性小,跨膜区且改变氨基酸性质的突变,可能影响蛋白的空间结构而改变膜通道的通透性。本文为进一步研究GJB2基因突变与聋病的关联性及分子发病机制提供了理论依据,同时,这种研究思路对其它疾病的相关研究具有一定的借鉴价值。     

EMS诱变水稻创制抗咪唑啉酮除草剂种质

为了创制水稻非转基因抗除草剂种质,丰富我国水稻轻简化栽培杂草防治所需的配套种质资源,本研究以0.5％(w/v)甲磺酸乙酯(EMS)水溶液诱变水稻种子14 h,然后以咪唑啉酮类除草剂筛选3 ～4叶期的M2水稻幼苗.结果发现,M2幼苗茎叶喷雾160 mg·L-1甲咪唑烟酸15 d后,非抗性苗黄化、生长被抑制甚至枯死,抗性苗...     

Protein biofortified sorghum: effect of processing into traditional african foods on their protein quality

Protein biofortification into crops is a means to combat childhood protein-energy malnutrition (PEM) in developing countries, by increasing the bioavailability of protein in staple plant foods and ensuring sustainability of the crop. Protein biofortification of sorghum has been achieved by both chemically induced mutation and genetic engineering. For this biofortification to be effective, the improved protein quality in the grain must be retained when it is processed into staple African foods. Suppression of kafirin synthesis by genetic engineering appeared to be superior to improved protein digestibility by chemical mutagenesis, because both the lysine content and protein digestibility were substantially improved and maintained in a range of African foods. For the genetically engineered sorghums, the protein digestibility corrected amino acid score was almost twice that of their null controls and considerably higher than the high protein digestibility sorghum type. Such protein biofortified sorghum has considerable potential to alleviate PEM.     

Regulation of lysine metabolism and endosperm protein synthesis by the opaque-5 and opaque-7 maize mutations

Two high lysine maize endosperm mutations, opaque-5 (o5) and opaque-7 (o7), were biochemically characterized for endosperm protein synthesis and lysine metabolism in immature seeds. Albumins, globulins, and glutelins, which have a high content of lysine, were shown to be increased in the mutants, whereas zeins, which contain trace concentrations of lysine, were reduced in relation to the wild-type lines B77xB79+ and B37+. These alterations in the storage protein fraction distribution possibly explain the increased concentration of lysine in the two mutants. Using two-dimensional polyacrylamide gel electrophoresis of proteins of mature grains, variable amounts of zein polypeptides were detected and considerable differences were noted between the four lines studied. The analysis of the enzymes involved in lysine metabolism indicated that both mutants have reduced lysine catabolism when compared to their respective wild types, thus allowing more lysine to be available for storage protein synthesis.