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Transgenic soybean plants overexpressing O-acetylserine sulfhydrylase accumulate enhanced levels of cysteine and Bowman–Birk protease inhibitor in seeds

机译:过表达O-乙酰丝氨酸巯基化酶的转基因大豆植物中种子中的半胱氨酸和Bowman–Birk蛋白酶抑制剂的含量增加

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Soybeans provide an excellent source of protein in animal feed. Soybean protein quality can be enhanced by increasing the concentration of sulfur-containing amino acids. Previous attempts to increase the concentration of sulfur-containing amino acids through the expression of heterologous proteins have met with limited success. Here, we report a successful strategy to increase the cysteine content of soybean seed through the overexpression of a key sulfur assimilatory enzyme. We have generated several transgenic soybean plants that overexpress a cytosolic isoform of O-acetylserine sulfhydrylase (OASS). These transgenic soybean plants exhibit a four- to tenfold increase in OASS activity when compared with non-transformed wild-type. The OASS activity in the transgenic soybeans was significantly higher at all the stages of seed development. Unlike the non-transformed soybean plants, there was no marked decrease in the OASS activity even at later stages of seed development. Overexpression of cytosolic OASS resulted in a 58–74% increase in protein-bound cysteine levels compared with non-transformed wild-type soybean seeds. A 22–32% increase in the free cysteine levels was also observed in transgenic soybeans overexpressing OASS. Furthermore, these transgenic soybean plants showed a marked increase in the accumulation of Bowman–Birk protease inhibitor, a cysteine-rich protein. The overall increase in soybean total cysteine content (both free and protein-bound) satisfies the recommended levels required for the optimal growth of monogastric animals.
机译:大豆为动物饲料提供了极好的蛋白质来源。增加含硫氨基酸的浓度可以提高大豆蛋白的品质。先前通过表达异源蛋白质来增加含硫氨基酸的浓度的尝试取得了有限的成功。在这里,我们报告了一种成功的策略,可以通过过量表达关键的硫同化酶来增加大豆种子中的半胱氨酸含量。我们已经产生了几种过表达O-乙酰丝氨酸巯基化酶(OASS)的胞质亚型的转基因大豆植物。与未转化的野生型植物相比,这些转基因大豆植物的OASS活性提高了4至10倍。在种子发育的所有阶段,转基因大豆中的OASS活性均显着较高。与未转化的大豆植物不同,即使在种子发育的后期,OASS活性也没有明显降低。与未转化的野生型大豆种子相比,胞质OASS的过表达导致蛋白质结合的半胱氨酸水平增加58-74%。在过表达OASS的转基因大豆中,游离半胱氨酸水平也增加了22–32%。此外,这些转基因大豆植物显示出Bowman–Birk蛋白酶抑制剂(一种富含半胱氨酸的蛋白质)的积累量显着增加。大豆总半胱氨酸含量的总体增加(游离和结合蛋白质的含量)均满足单胃动物最佳生长所需的推荐水平。

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