MALATE-DEHYDROGENASE AND GLUCOSE-6-PHOSPHATE-DEHYDROGENASE, KEY MARKERS FOR STUDYING THE GENETIC DIVERSITY OF ACTINOBACILLUS-ACTINOMYCETEMCOMITANS

Cell-free extracts of strains belonging to the 5 serotypes of A. actinomycetemcomitans were screened for several enzymes. Enzymes representative of the pentose phosphate pathway/hexose monophosphate shunt and the TCA cycle were present. Of these glucose-6-phosphate dehydrogenase (G6PDH) and malate dehydrogenase (MDH) were the most readily detected and stable. MDH and G6PDH retained more than 50% of their activities at alkaline pHs (10-11) for up to 6 h and 3 h at 25 degrees C, respectively, while at pH 6.5, 50% of their activities were lost within 2-3 h. The K-m for malate oxidation catalysed by MDH was 5.8x10(-4) M while that for glucose-6-phosphate oxidation was 2.0x10(-4) M. The pH optima for MDH and G6PDH oxidation activities were 10 and 9.5, respectively. Among the 5 designated serotypes of A. actinomycetemcomitans three groups were delineated by multilocus enzyme electrophoresis using MDH and G6PDH.