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首页> 外文期刊>Journal of the Science of Food and Agriculture >Purification of calmodulin from rice bran and activation of glutamate decarboxylase by Capo/calmodulin
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Purification of calmodulin from rice bran and activation of glutamate decarboxylase by Capo/calmodulin

机译：从米糠中纯化钙调蛋白并通过Capo /钙调蛋白激活谷氨酸脱羧酶

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BACKGROUND: d-Aminobutyric acid (GABA) is an important bioactive regulator, and its biosynthesis is primarily through the l-decarboxylation of glutamate by glutamate decarboxylase (GAD). In plants, it was verified that the production of GABA is regulated, in part, via Capo/calmodulin (CaM). Our preliminary studies showed that rice bran GAD is probably also a Capo/CaM dependent enzyme; hence, in the current investigation, we purified calmodulin from rice bran, and studied the effect of the Capo/calmodulin complex on the activity of rice bran GAD in vitro.RESULTS: CaM was purified to homogeneity from the rice bran by a combined protocol involving TCA precipitation, heat treatment, and hydrophobic interaction chromatography, with the purification fold and recovery of 851.7 and 55.6%, respectively. This protein had similar amino acid composition as the CaMs from other higher plants. The rice bran GAD was found to be quite sensitive to the Capo/CaM complex at pH 7.0, and addition of exogenous EGTA or TFP efficiently inhibited the stimulatory effect of Capo/CaM complex. At a separate concentration of Capo and CaM of 200 omol Lp# and 150 nmol Lp#, the rice bran GAD was significantly enhanced 3-fold. Moreover, upon binding Capo, CaM underwent a conformational change that facilitated a more obvious emergency of phenylalanine and tyrosine residues.CONCLUSION: This investigation provided preliminary information for the development of a GABA-based, cost-effective rice bran GAD-related functional food. Copyright
机译：背景：d-氨基丁酸（GABA）是重要的生物活性调节剂，其生物合成主要是通过谷氨酸脱羧酶（GAD）对谷氨酸进行l-脱羧。在植物中，已证实GABA的产生部分受Capo /钙调蛋白（CaM）的调节。我们的初步研究表明，米糠GAD可能也是Capo / CaM依赖的酶。因此，在目前的研究中，我们从米糠中纯化钙调蛋白，并研究了Capo /钙调蛋白复合物对米糠GAD活性的体外影响。 TCA沉淀法，热处理法和疏水作用色谱法的纯化倍数和回收率分别为851.7和55.6％。该蛋白质的氨基酸组成与其他高等植物的CaM相似。发现米糠GAD在pH 7.0下对Capo / CaM复合物非常敏感，添加外源EGTA或TFP有效抑制Capo / CaM复合物的刺激作用。在Capo和CaM的浓度分别为200 omol Lp＃和150 nmol Lp＃时，米糠GAD显着提高了3倍。此外，在结合Capo后，CaM发生了构象变化，促进了苯丙氨酸和酪氨酸残基的更明显的紧急情况。结论：这项研究为开发基于GABA的，具有成本效益的米糠GAD相关功能食品提供了初步信息。版权

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