Partial purification and anti-leukemic activity of L-asparaginase enzyme of the actinomycete strain LA-29 isolated from the estuarine fish, Mugil cephalus (Linn.).

The actinomycete strain LA-29 isolated from the gut contents of the fish, Mugil cephalus of the Vellar estuary showed excellent L-asparaginase activity The enzyme was purified 18-fold and the final recovery of protein was 1.9%, which exhibited an activity of 13.57 IU/mg protein. The partially purified L-asparaginase inhibited the growth of leukemia cells in male wistar rats. Average survival period of the rats was more in an optimum enzyme dose of 100 units and the survival period was less when the dosages were increased and at the same time the enzyme became less effective when the dosages were decreased. Higher survival of 17.2 days was recorded when 100 units of the enzyme was given in three intermittent doses (50/25/25 units) at the interval of 24 hr. Analysis of cell components of the strain LA-29 has revealed the wall type-I which is the characteristic of the genus Streptomyces. Further the morphological, physiological and biochemical features along with the micromorphological results obtained for the strain LA-29 were compared with that of the Streptomyces species found in Bergey's Manual of Determinative Bacteriology and the strain LA-29 has been tentatively identified as Streptomyces canus.

Studies on L-asparaginase enzyme of actinomycetes isolated from estuarine fishes.

Actinomycetes were isolated from different organs viz. skin, gills and gut contents of three species of fishes viz. Mugil cephalus (Linnaeus, 1758), Chanos chanos (Forskal, 1775) and Etroplus suratensis (Bloch, 1780) using three different media from the Vellar estuary, situated along the southeast coast of India. Among the three fishes, M. cephalus harboured highest number of actinomycetes population in all the three body parts examined followed by C. chanos and E. suratensis. Out of the three body parts of all fishes, gut contents had highest actinomycetes population followed by gills and skin. Among the three media used for isolation of actinomycetes, Kuster's agar medium was found to be suitable than the starch casein agar and glucose asparagine agar media. Out of the 40 strains isolated, only six strains (LA-2, LA-8, LA-15, LA-20, LA-29 and LA-35) showed significant L-asparagianse activity and were taken up for further studies. Impact of various physical and chemical factors such as pH, temperature, sodium chloride concentration, carbon sources and amino acids on the growth of actinomycetes and L-asparaginase activity was also studied. Optimum growth and enzyme activity was noticed under pH 7 to 8, temperature 37 degrees C, 1-2% sodium chloride concentration, sucrose as carbon source and without any amino acids. Analysis of the cell components of the isolated strains has revealed the wall type-I (the wall type-I is typical for the genus Streptomyces) and the strains were micromorphologically similar to the genus Streptomyces. Hence, the morphological, physiological and biochemical along with the micromorphological results obtained for the L-asparaginase producing strains were compared and the strains were tentatively identified as Streptomyces aureofasciculus (LA-2), S. chattanoogenesis (LA-8), S. hawaiiensis (LA-15), S. orientalis (LA-20), S. canus (LA-29) and S. olivoviridis (LA-35).