The Diversity of Polyketide Synthase Genes from Sugarcane-Derived Fungi

The chemical ecology and biotechnological potential of metabolites from endophytic and rhizosphere fungi are receiving much attention. A collection of 17 sugarcane-derived fungi were identified and assessed by PCR for the presence of polyketide synthase (PKS) genes. The fungi were all various genera of ascomycetes, the genomes of which encoded 36 putative PKS sequences, 26 shared sequence homology with beta-ketoacyl synthase domains, while 10 sequences showed homology to known fungal C-methyltransferase domains. A neighbour-joining phylogenetic analysis of the translated sequences could group the domains into previously established chemistry-based clades that represented non-reducing, partially reducing and highly reducing fungal PKSs. We observed that, in many cases, the membership of each clade also reflected the taxonomy of the fungal isolates. The functional assignment of the domains was further confirmed by in silico secondary and tertiary protein structure predictions. This genome mining study reveals, for the first time, the genetic potential of specific taxonomic groups of sugarcane-derived fungi to produce specific types of polyketides. Future work will focus on isolating these compounds with a view to understanding their chemical ecology and likely biotechnological potential.

Isolation and biochemical, functional and structural characterization of a novel L-amino acid oxidase from Lachesis muta snake venom

The aim of this study was the isolation of the LAAO from Lachesis muta venom (LmLAAO) and its biochemical, functional and structural characterization. Two different purification protocols were developed and both provided highly homogeneous and active LmLAAO. It is a homodimeric enzyme with molar mass around 120 kDa under non-reducing conditions, 60 kDa under reducing conditions in SDS-PAGE and 60852 Da by mass spectrometry. Forty amino acid residues were directly sequenced from LmLAAO and its complete cDNA was identified and characterized from an Expressed Sequence Tags data bank obtained from a venom gland. A model based on sequence homology was manually built in order to predict its three-dimensional structure. LmLAAO showed a catalytic preference for hydrophobic amino acids (K-m of 0.97 mmol/L with Leu). A mild myonecrosis was observed histologically in mice after injection of 100 mu g of LmLAAO and confirmed by a 15-fold increase in CK activity. LmLAAO induced cytotoxicity on AGS cell line (gastric adenocarcinoma, IC50: 22.7 mu g/mL) and on MCF-7 cell line (breast adenocarcinoma, IC50:1.41 mu g/mL). It presents antiparasitic activity on Leishmania brasiliensis (IC50: 2.22 mu g/nnL), but Trypanosoma cruzi was resistant to LmLAAO. In conclusion, LmLAAO showed low systemic toxicity but important in vitro pharmacological actions. (C) 2012 Elsevier Ltd. All rights reserved.

Homology in a context dependent predatory behavior in spiders (Araneae)

Stereotyped behaviors have been routinely used as characters for phylogeny inference, but the same cannot be said of the plastic aspects of performance, which routinely are taken as a result of ecological processes. In this paper we examine the evolution of one of these plastic behavioral phenotypes, thus fostering a bridge between ecological and evolutionary processes. Foraging behavior in spiders is context dependent in many aspects, since it varies with prey type and size, spider nutritional and developmental state, previous experience and, in webweavers, is dependent on the structure of the web. Reeling is a predatory tactic typical of cobweb weavers (Theridiidae), in which the spider moves the prey toward her by pulling the capture thread (gumfoot) to which it is adhered. Predatory reeling is dependent on the gumfoot for its expression, and has not been previously reported in orbweavers. In order to investigate the evolution of this web dependent behavior, we built artificial, pseudogumfoot lines in orbwebs and registered parameters of the predatory tactics in this modified web. Aspects of the predatory tactics of 240 individuals (12 species in 4 families) were measured, and the resulting data were optimized on the phylogeny of Orbiculariae. All species perform predatory reeling with the pseudogumfoot lines. Thus, predatory reeling is homologous for the whole Orbiculariae group. In nature, holes made by insects in ecribellate orbs produce pseudogumfoot lines (similar to out experimentally modified webs), and thus reeling occurred naturally in ecribellates. Nevertheless, outside lab conditions, predatory reeling does not occur among cribellate orbweavers, so that this behavior could not have been selected for in the cribellate ancester of orbweavers. Cribellate spiders are flexible enough as to present novel and adaptive predatory responses (reeling) even when exposed for the first time to conditions outside their usual environment. Thus, the evolution of reeling suggests and alternative mechanism for the production of evolutionary novelties; that is, the exploration of unusual ecological conditions and of the regular effects these abnormal conditions have on phenotype expression.