Pilot‐Scale Purification of Zeamatin, an Antifungal Protein from Maize

Abstract

Zeamatin is an antifungal protein isolated from the seeds of Zea mays. A practical process for isolation and purification was developed to increase recovery yields for future testing of zeamatin as a novel therapeutic drug. Zeamatin was extracted with buffer from corn meal milled to flour. Solids were removed from the extract using pressure filtration, and zeamatin was purified using two separate reverse-phase chromatography steps. Determination of the amino-terminal amino acid sequence, Western blotting using anti-zeamatin antibody, and activity against Candida albicans were used to confirm that the purified protein was zeamatin. From 5 kg of corn meal, approximately 110 mg of zeamatin was purified to apparent homogeneity.

Purification of Recombinant Green Fluorescent Protein Using Chromatofocusing with a pH Gradient Composed of Multiple Stepwise Fronts

“…The results of the present study can also be compared to those of a recent related study where two stages of reversed-phase chromatography, one of which was carried out in a low-pressure column while the other was carried out in a high-performance column, were used to purify zeamatin with corn seeds as a starting material (35). The overall process consisted of an initial extraction step, a capture step accomplished using low-pressure chromatography together with a stepwise solvent gradient, a membrane concentration step followed by a diafiltration step, a second chromatographic step accomplished using a high-performance column together with a stepwise solvent gradient, and a second series of membrane concentration and diafiltration steps.…”

Antifungal‐protein production in maize (Zea mays) suspension cultures

“…However, their use for the production of proteins of biological interest is not very common. For this, the cell‐culture system described in the present paper is very appealing from the biotechnological point of view, representing a valid alternative to the direct purification of zeamatin from seeds [27]. Moreover, the amounts of zeamatin could be enhanced by the use of appropriate elicitors such as jasmonic acid, salicylic acid or laminarin oligosaccharides [24,25].…”

Brassiparin, an antifungal peptide fromBrassica parachinensisseeds

“…Antifungal proteins and peptides have been isolated from a variety of organisms including plants (Wilson et al . 2000; Wang and Ng 2001, 2003; Fujimura et al .…”