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氮通过调节鲍曼-伯克蛋白酶抑制剂的积累来降低大豆(Glycine max [L.] Merr.)中含硫氨基酸的含量。

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

作者信息

Krishnan Hari B, Bennett John O, Kim Won-Seok, Krishnan Ammulu Hari, Mawhinney Thomas P

机构信息

Agricultural Research Service, U.S. Department of Agriculture, Columbia, Missouri 65211, USA.

出版信息

J Agric Food Chem. 2005 Aug 10;53(16):6347-54. doi: 10.1021/jf050510i.

Abstract

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.

摘要

大豆一般含有35%-40%的蛋白质。目前正在努力进一步提高这种蛋白质含量,从而提高其营养价值。尽管较高的蛋白质含量是一个理想的特性,但这种增加是否会伴随着蛋白质质量的提高尚不清楚。通过增加含硫氨基酸半胱氨酸和蛋氨酸的浓度,可以显著提高大豆蛋白质质量。为了确定蛋白质数量和质量之间是否存在相关性,对蛋白质含量不同的大豆的氨基酸进行了比较。蛋白质含量较高的大豆中,含硫氨基酸的百分比显著较低,而蛋白质含量较低的大豆中,半胱氨酸和蛋氨酸的含量较高。施氮提高了蛋白质含量,但降低了含硫氨基酸的含量。对低蛋白大豆种子薄片进行透射电子显微镜检查发现,有几个蛋白质储存液泡部分充满了储存蛋白。大豆种子蛋白的荧光二维差异凝胶电泳显示,施氮有利于β-伴大豆球蛋白β亚基的积累,同时减少了富含半胱氨酸的蛋白质鲍曼-伯克蛋白酶抑制剂(BBI)的积累。对60%异丙醇提取的蛋白质进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示,随着蛋白质含量的增加,BBI的积累急剧减少。Northern印迹分析表明,氮对BBI基因的表达有负面影响。我们的结果表明,总蛋白质与含硫氨基酸含量之间的负相关主要是由BBI的差异积累介导的。

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