Purification, Characterization, and Functional Analysis of Isocitrate Lyase Isoforms from Corn Scutella using Ion-exchange Chromatography

Saba Hadi, Zahraa B. Mohammed, Huda F. Ramadan

Abstract


Background: The work described herein investigates the properties of the enzyme isocitrate lyase, which was purified from corn plants using advanced techniques such as sulfate precipitation and ion exchange chromatography. The results indicated the presence of two forms of the enzyme, ICL1 and ICL2, which differed in their molecular weights and enzymatic activities. The optimum pH for ICL1 was found to be 7.5, while for ICL2, it was 6. Additionally, the effects of glycine and glycolate on the enzyme's activity were studied, revealing elevated activities at the optimal concentrations of these substances.

Methods: The isocitrate lyase enzyme was purified from corn plants using ion exchange chromatography and sulfate precipitation techniques. Enzyme activity was assessed using spectrophotometric methods, and the molecular weight was determined through gel chromatography. Studies were conducted to investigate the influence of pH, glycine, and glycolate on enzyme activity.

Results: Two isoforms of isocitrate lyase, ICL1 and ICL2, were purified, exhibiting molecular masses of 164 kDa and 208 kDa, respectively. ICL1 demonstrated optimum activity at pH 7.5, while ICL2 exhibited optimum activity at pH 6. In this study, specific concentrations of glycine and glycolate were found to enhance the enzymatic activities of both isoforms.

Conclusion: This research provides significant insights into the characteristics of the isocitrate lyase enzyme in corn plants. The data indicate the presence of distinct enzyme forms with specific interactions in varying environmental conditions, which may be applicable in agricultural practices to increase crop yield and improve the metabolic turnover of organic acids.

Keywords: Isocitrate lyase isoenzymes; Corn scutella; Chromatography; Glycine; Glycolate


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DOI: http://dx.doi.org/10.62940/als.v12i2.2562

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