Wallace W
Department of Agricultural Biochemistry, Waite Agricultural Research Institute, University of Adelaide, Glen Osmond, South Australia, 5064 Australia.
Plant Physiol. 1973 Sep;52(3):197-201. doi: 10.1104/pp.52.3.197.
The nitrate reductase in the mature root extract of 3-day maize (Zea mays) seedlings was relatively labile in vitro. Insoluble polyvinylpyrrolidone used in the extraction medium produced only a slight increase in the stability of the enzyme. Mixing the mature root extract with that of the root tip promoted the inactivation of nitrate reductase in the latter. The inactivating factor in the mature root was separated from nitrate reductase by (NH(4))(2)SO(4) precipitation. Nitrate reductase was found in the 40% (NH(4))(2)SO(4) precipitate, while the inactivating factor was largely precipitated by 40 to 55% (NH(4))(2)SO(4). The latter fraction of the mature root inactivated the nitrate reductase isolated from the root tip, mature root, and scutellum. The inactivating factor, which has a Q(10) 15 to 25 C of 2.2, was heat labile, and hence has been designated as a nitrate reductase inactivating enzyme. The reduced flavin mononucleotide nitrate reductase was also inactivated, while an NADH cytochrome c reductase in nitrate-grown seedlings was inactivated but at a slower rate. The inactivating enzyme had no influence on the activity of nitrite reductase, glutamate dehydrogenase, xanthine oxidase, and isocitrate lyase. The activity of the nitrate reductase inactivating enzyme was not influenced by nitrate and was also found in the mature root of minus nitrate-grown seedlings.
3日龄玉米(Zea mays)幼苗成熟根提取物中的硝酸还原酶在体外相对不稳定。提取介质中使用的不溶性聚乙烯吡咯烷酮仅使该酶的稳定性略有增加。将成熟根提取物与根尖提取物混合会促进后者中硝酸还原酶的失活。通过硫酸铵沉淀将成熟根中的失活因子与硝酸还原酶分离。硝酸还原酶存在于40%硫酸铵沉淀中,而失活因子大部分在40%至55%硫酸铵中沉淀。成熟根的后一部分使从根尖、成熟根和盾片中分离出的硝酸还原酶失活。失活因子在15至25℃时的Q10为2.2,对热不稳定,因此被命名为硝酸还原酶失活酶。还原型黄素单核苷酸硝酸还原酶也被失活,而硝酸盐培养幼苗中的NADH细胞色素c还原酶被失活,但速度较慢。失活酶对亚硝酸还原酶、谷氨酸脱氢酶、黄嘌呤氧化酶和异柠檬酸裂解酶的活性没有影响。硝酸还原酶失活酶的活性不受硝酸盐影响,在无硝酸盐培养幼苗的成熟根中也有发现。