2 CLASS-I ALDOLASES IN KLEBSORMIDIUM-FLACCIDUM (CHAROPHYCEAE) - AN EVOLUTIONARY LINK FROM CHLOROPHYTES TO HIGHER-PLANTS

Two fructose‐bisphosphate aldolases(EC 4.1.2.13) from Klebsormidium flaccidum Silver, Mattox and Black‐well were purified by affinity elution from phosphocellulose. The two enzymes were subsequently separated by HPLC on an anion‐exchange column (QAE‐silica). The aldolase eluting first represented 5% of the total activity; the other aldolase represented the remaining activity. The activity of the enzymes was not reduced by the presence of 1 mM EDTA or increased by 0.1 mM Zn2+, establishing their character as class I type (Me2+ independent) aldolases. The Km(fructose‐1,6‐bisphosphate) values were 1.7 and 34.7 μM for the enzyme eluting first and second, respectively, from the QAE‐silica column. The subunit molecular masses, as determined by SDS‐PACE, were 40.5 and 37 kD; the specific activities of the purified enzymes were 7.9 and 24.7 · mg−1 protein, respectively. The two aldolases of K. flaccidum are homologous to the cytosol and chloroplast specific isoenzymes of higher plants by several criteria and are therefore probably located in the same cellular compartments in K. flaccidum. The Km and specific activity for the chloroplast aldolase of K. flaccidum are three times higher than for the chloroplast aldolase of higher plants, a remarkable difference. Immunotitration with specific antisera against the chloroplast aldolase of Chlamydomonas reinhardtii Dangeard and spinach showed that the chloroplast aldolase of K. flaccidum was immunochemically intermediate in structure to the respective aldolases of C. reinhardtii and higher plants. K. flaccidum is the second species of Charophyceae (besides Chara foetida Braun) with two class I aldolases as in higher plants whereas two species of Chlorophyceae have only one class I aldolase and, under some conditions, an additional class II (Me2+ dependent) aldolase. Thus, aldolases may turn out, in addition to the known enzymes of glycolate conversion and urea degradation, be a novel enzyme system to evaluate algal evolution along with cytological features. Copyright © 1990, Wiley Blackwell. All rights reserved