Purification and Characterization of Malic Enzyme of Entamoeba invadens: Evidence for Isoenzymes*

SYNOPSIS. Malic enzyme (EC 1.1.1.40) of Entamoeba invadens seems to be a single species of protein on zone-sedimentation, gel filtration, and gel electrophoresis. The molecular weight of the enzyme was estimated to be 120,000 and sedimentation coefficient close to 6S. Isoelectric focusing, however, showed malic enzyme to be composed of three isoenzymes of pI 5.8, 6.0, and 6.3. A procedure is described that yields an electrophoretically pure sample of each of the isoenzymes suitable for physical and chemical studies.     

A preliminary molecular phylogenetic assessment of the lichen moths (Lepidoptera: Erebidae: Arctiinae: Lithosiini) with comments on palatability and chemical sequestration

The lichen moth tribe Lithosiini is best known for its hypothesized larval feeding behaviour, lichenivory. The larvae of some species have been found to be capable of sequestering polyphenolics, and the adults of some species are unpalatable to vertebrate predators. However, the chemical basis for the defence is unknown. Here we reconstruct a phylogenetic hypothesis using likelihood methods (maximum likelihood, Bayesian inference) for 65 species of Lithosiini representing 37 genera and seven outgroup species using 2806 bp of sequence data obtained from two mitochondrial gene fragments (COI barcoding region, CytB), and two nuclear gene fragments (RpS5 and ribosomal gene region 28S). Lithosiine species representing four of the seven subtribes and unplaced taxa are included in the analysis. The deeper relationships within the tribe are not strongly supported, and the monophyly of three of the four subtribes is not supported. The placement of the fourth subtribe, Acsalina, is weakly supported. We also conduct a survey of the secondary metabolites present within adults of five species. The species are examined for the presence of lichen polyphenolics and plant secondary metabolites sequestered by arctiines. Seven lichen polyphenolics are identified in the taxa examined, but no plant secondary metabolites are found using either analytical method. The results of this study and prior reports in the literature of chemical sequestration and unpalatability are plotted onto the resulting Bayesian inference phylogeny to examine the evolution of chemical defence within Lithosiini. Species that sequester lichen polyphenolics and unpalatable species occur in each of the major clades recovered in the analysis.