Comparison of the phytohaemagglutinin from red kidney bean ( Phaseolus vulgaris) purified by different affinity chromatography

• Published in: Food chemistry Vol. 108; no. 1; pp. 394 - 401
• Main Authors: Ren, Jiaoyan, Shi, John, Kakuda, Yukio, Kim, Daniel, Xue, Sophia Jun, Zhao, Mouming, Jiang, Yueming, Sun, Jian
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.05.2008; [Amsterdam]: Elsevier Science; Elsevier
• Subjects: Affinity chromatography; Biological and medical sciences; Circular dichroism; Food industries; Fruit and vegetable industries; Fundamental and applied biological sciences. Psychology; Haemagglutinating; kidney beans; lectins; Phaseolus vulgaris; Phytohaemagglutinin; phytohemagglutinin; purification; Red kidney bean; red kidney beans; CD; PHA-L; AC; Tg; PHA-P; Affinity chromatography; Haemagglutinating; PHA; PHA-E; Red kidney bean; Phytohaemagglutinin; Circular dichroism; Vegetables; Bean; Red; Leguminosae; Phaseolus vulgaris; Dicotyledones; Angiospermae; Spermatophyta
• ISSN: 0308-8146; 1873-7072
• DOI: 10.1016/j.foodchem.2007.10.071

Affinity chromatography (AC) on Affi-Gel blue gel column and thyroglobulin (Tg)-Sepharose 4B column, respectively, were compared for their efficiency in purifying phytohaemagglutinin (PHA) from red kidney beans ( Phaseolus vulgaris). Considering the purity and haemagglutinating activity of the obtained samples, Affi-Gel blue gel exhibited less affinity for PHA than Tg-Sepharose matrix. Affi-Gel blue purified sample showed multiple bands in SDS-PAGE gel, which further confirmed that Affi-Gel blue bound non-PHA proteins as well as PHA. PHA purified by one-step Tg-Sepharose column gave significantly ( p < 0.05) higher purity (0.75 ± 0.13 mg PHA/mg lyophilized powder) than the sample purified by two-step (Affi-Gel blue first and then Tg-Sepharose) purification (0.62 ± 0.20 mg PHA/mg lyophilized powder). Circular dichroism (CD) spectra showed that the sample purified by one-step Tg-Sepharose column had similar secondary structures with the sample purified by two-step purification. Thus, one-step Tg-Sepharose purification was effective and time-saving for the preparation of PHA and a promising substitute for the two-step purification method.