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首页> 美国卫生研究院文献>Plant Physiology >Extraction and Partial Characterization of the Glycine Decarboxylase Multienzyme Complex from Pea Leaf Mitochondria
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Extraction and Partial Characterization of the Glycine Decarboxylase Multienzyme Complex from Pea Leaf Mitochondria

机译：豌豆线粒体中甘氨酸脱羧酶多酶复合物的提取及部分表征

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摘要

Glycine decarboxylase has been successfully solubilized from pea (Pisum sativum) leaf mitochondria as an acetone powder. The enzyme was dependent on added dithiothreitol and pyridoxal phosphate for maximal activity. The enzyme preparation could catalyze the exchange of CO2 into the carboxyl carbon of glycine, the reverse of the glycine decarboxylase reaction by converting serine, NH4+, and CO2 into glycine, and 14CO2 release from [1-14C]glycine. The half-maximal concentrations for the glycine-bicarbonate exchange reaction were 1.7 millimolar glycine, 16 millimolar NaH14CO2, and 0.006 millimolar pyridoxal phosphate. The enzyme (glycine-bicarbonate exchange reaction) was active in the assay conditions for 1 hour and could be stored for over 1 month. The enzymic mechanism appeared similar to that reported for the enzyme from animals and bacteria but some quantitative differences were noted. These included the tenacity of binding to the mitochondrial membrane, the concentration of pyridoxal phosphate needed for maximum activity, the requirement for dithiothreitol for maximum activity, and the total amount of activity present. Now that this enzyme has been solubilized, a more detailed understanding of this important step in photorespiration should be possible.
机译：甘氨酸脱羧酶已成功地从豌豆（Pisum sativum）叶线粒体中溶解为丙酮粉。该酶依赖于添加的二硫苏糖醇和磷酸吡al醛的最大活性。该酶制剂可通过将丝氨酸，NH4 + 和CO2转化为甘氨酸和 14 14 C]甘氨酸释放> CO2。甘氨酸-碳酸氢盐交换反应的半最大浓度为1.7毫摩尔甘氨酸，16毫摩尔NaH 14 CO2和0.006毫摩尔吡ido醛磷酸盐。该酶（甘氨酸-碳酸氢根交换反应）在测定条件下具有1小时的活性，可以保存1个月以上。酶学机制似乎与动物和细菌中酶的报道机制相似，但注意到一些定量差异。这些因素包括与线粒体膜结合的强度，最大活性所需的磷酸吡ido醛的浓度，最大活性所需的二硫苏糖醇以及存在的总活性。既然已经溶解了这种酶，那么应该有可能对光呼吸这一重要步骤有更详细的了解。

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