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. 1983 Jan;71(1):205–207. doi: 10.1104/pp.71.1.205

Purification of Squash NADH:Nitrate Reductase by Zinc Chelate Affinity Chromatography 1

Margaret G Redinbaugh 1,2, Wilbur H Campbell 1,2
PMCID: PMC1067205  PMID: 16662789

Abstract

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 Zn2+ 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.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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