ABSTRACT
alpha-Amylases from Drosophila virilis and D. repleta were partially purified by ion exchange chromatography. The two amylases share common characteristics for pH and cations effects, although with slight differences. D. virilis has optimal activity at pH 6.6 and D. repleta at pH 7.2. Calcium, sodium, and potassium cations activate amylolytic activity in both species but Ba2+ has an activation effect in D. repleta only. In contrast, there are major differences in thermal offbility and kinetics among amylases of the two species. D. virilis amylase is much more stable at high temperature and the optimal temperatures are very different between the two species, respectively, 45 degrees C and 30 degrees C for D. virilis and D. repleta. alpha-Amylase activity using different substrates is greater on starch than on glycogen in both species and still higher on amylose for D. virilis, the nonfungus feeder species. alpha-Amylase of D. repleta, the mycophagous species, has a better affinity to amylopectin and glycogen. Such differences in substrate specificity suggest adaptation to different resources in these species living in different habitats. Metabolic evolution seems to have occurred through a "tradeoff" between kinetic effectiveness and the nature of substrate, with a higher Vmax on amylose for D. virilis and a lower K(m) on glycogen for D. repleta.
Subject(s)
Drosophila/genetics , Ecosystem , Evolution, Molecular , Isoenzymes/genetics , alpha-Amylases/genetics , Adaptation, Physiological , Animals , Cations/pharmacology , Drosophila/metabolism , Hydrogen-Ion Concentration , Isoenzymes/metabolism , Kinetics , Species Specificity , Substrate Specificity , Temperature , alpha-Amylases/metabolismABSTRACT
Four xylanases were purified, two from the termite Macrotermes bellicosus workers (XIT and X2T) and two from its symbiotic fungus Termitomyces sp. (X1Mc and X2Mc). The analysis of the step required for the purification of X1T and X1Mc and the comparison of their different properties suggested that xylanases X1T and X1Mc were the same enzyme, X1. The determination of the reducing sugars by TLC revealed that X1 was an endoxylanase (EC 3.2.1.8) and X2T and X2Mc were endoxylanases (EC 3.2.1.37). The apparent molecular weights of the three xylanases, determined by SDS-polyacrylamide gel electrophoresis, were 36 kDa for X1, 56 kDa for X2T and 22.5 kDa for X2Mc. The optimal pH of the three xylanases was approximately 5.5, and Km values determined with birchwood xylan as substrate were 0.2% for X1, 0.1% for X2T and 0.3% for X2Mc, showing a high affinity for this substrate. The three enzymes differed also by their thermal stability.
