The effects of protein characteristics and the proportion of gluten on end-use quality in 13 Korean wheat cultivars for three years were verified in this study. Year, cultivar, and the interaction between the year and...The effects of protein characteristics and the proportion of gluten on end-use quality in 13 Korean wheat cultivars for three years were verified in this study. Year, cultivar, and the interaction between the year and the cultivar influenced protein characteristics, the proportion of gluten except for γ-and ω-gliadin using RP-HPLC(reversed-phase high-performance liquid chromatography), and end-use quality. Protein characteristics and the proportion of gluten in Korean wheat cultivars were between those of Australian standard white(ASW) and hard wheat(AH). Korean wheat cultivars exhibited a higher average α+β gliadin proportion than imported wheat, a γ-gliadin proportion similar to that of dark northern spring wheat, and the same ω-gliadin proportion as AH. They showed a bread-loaf volume intermediate between those of ASW and AH and a texture of cooked noodles similar to that of soft white wheat, but less springiness than imported wheat. The cookie diameter of Korean wheat cultivars was similar to that of hard red winter wheat. There was a correlation between bread-loaf volume and protein characteristics, except for the protein content in Korean wheat cultivars. Springiness and cohesiveness of cooked noodles were not correlated with protein characteristics, while hardness was correlated with the protein content and water absorption of a mixograph. Cookie diameter was negatively correlated with the sodium dodecyl sulfate(SDS) sedimentation volume and water absorption of a mixograph. The end-use quality was not correlated with any proportion of gluten composition. Principal component analysis(PCA) showed that the proportion of gluten was not related to the quality of the bread(both PCs, 81.3%), noodle(77.7%), and cookie(82.4%). PCA explained that Keumkang is suitable for superior bread, while Uri is good for cooked noodles and cookies.展开更多
ω-Secalin was an important factor influencing processing quality of wheat 1BL/1RS translocations. On the basis of four ω-secalin gene sequences cloned from Lankao 906 （a wheat cultivar with 1BL/1RS translocation） ...ω-Secalin was an important factor influencing processing quality of wheat 1BL/1RS translocations. On the basis of four ω-secalin gene sequences cloned from Lankao 906 （a wheat cultivar with 1BL/1RS translocation） with putative transcription activity, a pair of primers with suitable restriction endonucleases added at their 5＂ ends were designed to amplify the mature protein-coding regions of the four genes. After digestion with restriction endonucleases, the amplified products were ligated into the prokaryotic expression vector pET30a（＋）. The prokaryotically expressed recombinant proteins and gliadin isolated from the Lankao 906 seed were analyzed on the same acid polyacrylamide gel electrophoresis. All four prokaryotically expressed secalin bands had corresponding seed-expressed gliadin bands. The four corresponding gliadin bands were confirmed to be the expression products of the four ω-secalin genes by liquid chromatography tandem mass spectrometry （LC-MS/MS）. This conclusion was further confirmed because the four ω-secalin bands could be detected in all 14 wheat 1BL/1 RS translocation cultivars used in the study, although there was some interference for the detection of one ω-secalin band from nearby wheat gliadin bands. The sequence information of ω-secalin genes with expression activity will be helpful for improving the processing quality of wheat with 1BL/1RS translocations by using RNA interference method to silence the expression of the ω-secalin genes.展开更多
分析确定大豆贮藏蛋白的种类、序列同源性、进化关系、理化特征和空间结构。以大豆(Glycine max Wm82.a2.v1)基因组为材料,采用数据库在线工具和生物信息学软件对大豆贮藏蛋白基因的染色体定位,蛋白序列信息进行分析预测,并对其3-D结构...分析确定大豆贮藏蛋白的种类、序列同源性、进化关系、理化特征和空间结构。以大豆(Glycine max Wm82.a2.v1)基因组为材料,采用数据库在线工具和生物信息学软件对大豆贮藏蛋白基因的染色体定位,蛋白序列信息进行分析预测,并对其3-D结构进行同源模建。大豆贮藏蛋白基因具有多拷贝和单基因多种转录拼接方式并存的遗传模式,其蛋白序列相似度较高,系统进化树分析表明其聚为5支,对应5种类型蛋白质。除7S碱性球蛋白外,其他贮藏蛋白的理化性质接近,含有较高比例的鲜味氨基酸。大豆贮藏蛋白空间结构各异,11S球蛋白、7Sβ-伴球蛋白和碱性球蛋白分别以六聚体、三聚体和四聚体形式存在,2S清蛋白表空间结构松散,由α-螺旋和无规卷曲构成。该研究可为大豆贮藏蛋白的深入研究以及大豆加工食品、大豆酿造调味品的开发提供理论参考。展开更多
The occurrence of twin-arginine motifs （-R-R-） in the amino acid sequences of animal pro-proteins frequently defines the cleavage site（s） for their structural/functional maturation. No information is available on ...The occurrence of twin-arginine motifs （-R-R-） in the amino acid sequences of animal pro-proteins frequently defines the cleavage site（s） for their structural/functional maturation. No information is available on the presence and possible biological meaning of these motifs in the seed storage proteins. In this work, a novel endopeptidase activity with cleavage specificity to twin-arginine pairs has been detected in mature dry Lupinus albus seeds. The endopeptidase was tested with a number of endogenous and exogenous protein substrates, which were selected according to the pres- ence of one or more twin-arginine residue motifs in their amino acid sequences. The observed hydrolysis patterns were limited and highly specific. Partial proteolysis led to stable polypeptide fragments that were characterized by 1- and 2-D electrophoresis. Selected polypeptides were submitted to N-terminal amino acid sequencing and mass spectrometry anal- yses, These approaches, supported by bioinformatic analysis of the available sequences, allowed the conclusion that the polypeptide cleavage events had occurred at the peptide bonds comprised between twin-arginine residue pairs with all tested protein substrates. The endopeptidase activity was inhibited by 4-（2-AminoEthyl）Benzene-Sulphonyl Fluoride hy- drochloride （AEBSF）, leupeptin, and serine proteinase protein inhibitors, while it was not affected by pepstatin, trans- EpoxysuccinyI-L-leucylamido（4-guanidino）butane （E64）, and ethylenediaminetetraacetic acid （EDTA）, thus qualifying the Arg-Arg cleaving enzyme as a serine endopeptidase. The structural features of storage proteins from lupin and other legume seeds strongly support the hypothesis that the occurrence of an endopeptidase activity cleaving -R-R- bonds may be functional to facilitate their degradation at germination and possibly generate polypeptide fragments with specific biological activity.展开更多
基金the support of Cooperative Research Program for Agriculture Science & Technology Development (Project title: Establishment of quality criteria for high uniformity in end-use of Korean wheat cultivars, PJ011009), Rural Development Administration, Republic of Korea
文摘The effects of protein characteristics and the proportion of gluten on end-use quality in 13 Korean wheat cultivars for three years were verified in this study. Year, cultivar, and the interaction between the year and the cultivar influenced protein characteristics, the proportion of gluten except for γ-and ω-gliadin using RP-HPLC(reversed-phase high-performance liquid chromatography), and end-use quality. Protein characteristics and the proportion of gluten in Korean wheat cultivars were between those of Australian standard white(ASW) and hard wheat(AH). Korean wheat cultivars exhibited a higher average α+β gliadin proportion than imported wheat, a γ-gliadin proportion similar to that of dark northern spring wheat, and the same ω-gliadin proportion as AH. They showed a bread-loaf volume intermediate between those of ASW and AH and a texture of cooked noodles similar to that of soft white wheat, but less springiness than imported wheat. The cookie diameter of Korean wheat cultivars was similar to that of hard red winter wheat. There was a correlation between bread-loaf volume and protein characteristics, except for the protein content in Korean wheat cultivars. Springiness and cohesiveness of cooked noodles were not correlated with protein characteristics, while hardness was correlated with the protein content and water absorption of a mixograph. Cookie diameter was negatively correlated with the sodium dodecyl sulfate(SDS) sedimentation volume and water absorption of a mixograph. The end-use quality was not correlated with any proportion of gluten composition. Principal component analysis(PCA) showed that the proportion of gluten was not related to the quality of the bread(both PCs, 81.3%), noodle(77.7%), and cookie(82.4%). PCA explained that Keumkang is suitable for superior bread, while Uri is good for cooked noodles and cookies.
基金supported by the Natural Science Foundation of Hebei Province,China (C2014301005)
文摘ω-Secalin was an important factor influencing processing quality of wheat 1BL/1RS translocations. On the basis of four ω-secalin gene sequences cloned from Lankao 906 （a wheat cultivar with 1BL/1RS translocation） with putative transcription activity, a pair of primers with suitable restriction endonucleases added at their 5＂ ends were designed to amplify the mature protein-coding regions of the four genes. After digestion with restriction endonucleases, the amplified products were ligated into the prokaryotic expression vector pET30a（＋）. The prokaryotically expressed recombinant proteins and gliadin isolated from the Lankao 906 seed were analyzed on the same acid polyacrylamide gel electrophoresis. All four prokaryotically expressed secalin bands had corresponding seed-expressed gliadin bands. The four corresponding gliadin bands were confirmed to be the expression products of the four ω-secalin genes by liquid chromatography tandem mass spectrometry （LC-MS/MS）. This conclusion was further confirmed because the four ω-secalin bands could be detected in all 14 wheat 1BL/1 RS translocation cultivars used in the study, although there was some interference for the detection of one ω-secalin band from nearby wheat gliadin bands. The sequence information of ω-secalin genes with expression activity will be helpful for improving the processing quality of wheat with 1BL/1RS translocations by using RNA interference method to silence the expression of the ω-secalin genes.
文摘分析确定大豆贮藏蛋白的种类、序列同源性、进化关系、理化特征和空间结构。以大豆(Glycine max Wm82.a2.v1)基因组为材料,采用数据库在线工具和生物信息学软件对大豆贮藏蛋白基因的染色体定位,蛋白序列信息进行分析预测,并对其3-D结构进行同源模建。大豆贮藏蛋白基因具有多拷贝和单基因多种转录拼接方式并存的遗传模式,其蛋白序列相似度较高,系统进化树分析表明其聚为5支,对应5种类型蛋白质。除7S碱性球蛋白外,其他贮藏蛋白的理化性质接近,含有较高比例的鲜味氨基酸。大豆贮藏蛋白空间结构各异,11S球蛋白、7Sβ-伴球蛋白和碱性球蛋白分别以六聚体、三聚体和四聚体形式存在,2S清蛋白表空间结构松散,由α-螺旋和无规卷曲构成。该研究可为大豆贮藏蛋白的深入研究以及大豆加工食品、大豆酿造调味品的开发提供理论参考。
文摘The occurrence of twin-arginine motifs （-R-R-） in the amino acid sequences of animal pro-proteins frequently defines the cleavage site（s） for their structural/functional maturation. No information is available on the presence and possible biological meaning of these motifs in the seed storage proteins. In this work, a novel endopeptidase activity with cleavage specificity to twin-arginine pairs has been detected in mature dry Lupinus albus seeds. The endopeptidase was tested with a number of endogenous and exogenous protein substrates, which were selected according to the pres- ence of one or more twin-arginine residue motifs in their amino acid sequences. The observed hydrolysis patterns were limited and highly specific. Partial proteolysis led to stable polypeptide fragments that were characterized by 1- and 2-D electrophoresis. Selected polypeptides were submitted to N-terminal amino acid sequencing and mass spectrometry anal- yses, These approaches, supported by bioinformatic analysis of the available sequences, allowed the conclusion that the polypeptide cleavage events had occurred at the peptide bonds comprised between twin-arginine residue pairs with all tested protein substrates. The endopeptidase activity was inhibited by 4-（2-AminoEthyl）Benzene-Sulphonyl Fluoride hy- drochloride （AEBSF）, leupeptin, and serine proteinase protein inhibitors, while it was not affected by pepstatin, trans- EpoxysuccinyI-L-leucylamido（4-guanidino）butane （E64）, and ethylenediaminetetraacetic acid （EDTA）, thus qualifying the Arg-Arg cleaving enzyme as a serine endopeptidase. The structural features of storage proteins from lupin and other legume seeds strongly support the hypothesis that the occurrence of an endopeptidase activity cleaving -R-R- bonds may be functional to facilitate their degradation at germination and possibly generate polypeptide fragments with specific biological activity.
