{"title":"Spinach nitrate reductase: Further purification and removal of ‘nicked’ sub-units by affinity chromatography","authors":"R.J. Fido, B.A. Notton","doi":"10.1016/0304-4211(84)90208-6","DOIUrl":null,"url":null,"abstract":"<div><p>NADH-nitrate reductase (EC 1.6.6.1) from spinach (<em>Spinacea oleracea</em> L. v. Noorman) has been purified 3400-fold by a multi-stage procedure involving streptomycin sulphate, (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>, hydroxylapatite, molecular seiving and two stages of affinity chromatography using blue-Sepharose and 5′ AMP-Sepharose. The enzyme has a specific activity of 51 μmol NO<sub>2</sub><sup>−</sup> produced min<sup>−1</sup> mg<sup>−1</sup> and a functional haem with extinction coefficients (mM) of 127 at 412 nm (oxidized) and 172 at 423 (NADH-reduced). Gel electrophoresis indicates two sub-units of approx. 110 000 and 120 000.</p></div>","PeriodicalId":20221,"journal":{"name":"Plant Science Letters","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1984-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0304-4211(84)90208-6","citationCount":"23","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Plant Science Letters","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0304421184902086","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 23
Abstract
NADH-nitrate reductase (EC 1.6.6.1) from spinach (Spinacea oleracea L. v. Noorman) has been purified 3400-fold by a multi-stage procedure involving streptomycin sulphate, (NH4)2SO4, hydroxylapatite, molecular seiving and two stages of affinity chromatography using blue-Sepharose and 5′ AMP-Sepharose. The enzyme has a specific activity of 51 μmol NO2− produced min−1 mg−1 and a functional haem with extinction coefficients (mM) of 127 at 412 nm (oxidized) and 172 at 423 (NADH-reduced). Gel electrophoresis indicates two sub-units of approx. 110 000 and 120 000.
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