Effect of assay conditions on the measurement of dehydrogenase activity of Streptomyces venezuelae using triphenyl tetrazolium chloride

Jadomycin is an antibiotic that has shown activities against bacteria, yeasts and fungi as well as cytotoxic properties to cancer cells. Because of the wide range of its inhibitory actions, jadomycin shows promise as a novel antibiotic and cancer treatment drug. Streptomyces venezuelae are aerobic bacteria that produce jadomycin and the size of bacterial population can significantly affect the yield of jadomycin. Therefore, the bacterial population must be accurately measured in order to standardize the reproducibility of jadomycin production process. In this study, a dehydrogenase activity measurement test, using triphenyl tetrazolium chloride (TTC), was used to measure the dehydrogenase activity of Streptomyces venezuelae during growth in maltose-yeast extract-malt extract (MYM) broth. The aims were to evaluate the effectiveness of the test for measuring microbial growth and to study the effects of the test conditions (incubation time, incubation temperature and medium pH) on triphenyl formazan (TF) yield. The results showed that the TF yield was highly correlated to the optical density. The highest TF yield was observed at a pH of 6 at all incubation times and temperature. Lower TF yields were obtained at higher temperature (40 and 50oC) compared to those obtained at lower temperatures (22 and 30oC). The difference between the yields obtained at 22oC and 30oC were not significant. The differences between incubation time were also not significant. The recommended test conditions are an incubation time of 1 hour at a temperature of 30oC and a pH of 6 followed by three extractions using methanol.