Purification of Squash NADH:Nitrate Reductase by Zinc Chelate Affinity Chromatography | Zendy

Margaret G. Redinbaugh | Zendy; Wilbur Campbell | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

Purification of Squash NADH:Nitrate Reductase by Zinc Chelate Affinity Chromatography

Author(s) -

Margaret G. Redinbaugh,

Wilbur Campbell

Publication year - 1983

Publication title -

plant physiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 3.554

H-Index - 312

eISSN - 1532-2548

pISSN - 0032-0889

DOI - 10.1104/pp.71.1.205

Subject(s) - nitrate reductase , cucurbita maxima , chromatography , chemistry , elution , squash , nitrate , affinity chromatography , enzyme , chromatofocusing , reductase , zinc , chelation , biochemistry , specific activity , isoelectric point , biology , botany , inorganic chemistry , organic chemistry

NADH:nitrate reductase (EC 1.6.6.1) was isolated and purified from the green cotyledons of 5-day-old squash seedlings (Cucurbita maxima L.). The 10-hour purification procedure consisted of two steps: direct application of crude enzyme to blue Sepharose and specific elution with NADH followed by direct application of this effluent to a Zn(2+) column with elution by decreasing the pH of the phosphate buffer from 7.0 to 6.2. The high specific activity (100 micromoles per minute per milligram protein) and high recovery (15-25%) of electrophoretically homogeneous nitrate reductase show that the enzyme was not damaged by exposure to the bound zinc. With this procedure, homogeneous nitrate reductase can be obtained in yields of 0.5 milligram per kilogram cotyledons.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research