Cloning and expression of a Hemocyanin isolated from the centipede Cryptops iheringi

Among the centipede of the chilopode Myriapoda class, the Cryptops genus is one of the most associated with accidents in humans in the metropolitan region of the state of São Paulo. To date, there is no study in the literature about Cryptops iheringi toxins. Thus, in this work a transcriptomic analysis of the C. iheringi venom gland was performed to obtain a profile of the toxins of this species. In addition, the crude venom was subjected to mass spectrometry analysis to establish an association between unknown sequences. These approaches for the construction of a general profile of the venom gland expression of this species led to the he identification of a Hemocyanin (Hc) subunit.

Hemocyanins are copper-containing, respiratory proteins that occur in the haemolymph of many arthropod species. Here, we report the presence of Hc in the chilopode Myriapoda C. iheringi. Such respiratory proteins have long been considered unnecessary in Myriapoda, due to its tracheal systems.

These respiratory proteins are potent immunogens, which induce the synthesis of large amounts of specific antibodies. Studies pointed out its interaction with polymorphonuclear monocytes and lymphocytes and in vitro tests have shown a potential anticancer activit, with in vitro significant inhibition of the growth of cancerous strains of the breast, pancreas and prostate. Currently scientific data is mostly limited to the study of native Hc of M. crenulata molluscs, therefore, the biotechnological potential of Hcs isolated from centipedes is still unexplored.

Herein, the Hc sequence that was present in both proteome and transcriptome analysis have signal peptide and 76 kDa range. The Hc subunit sequence was synthesized with codon optimization for bacteria expression and the protein expressed as inclusion bodies. Refolding attempts provided soluble forms of Hc. At the moment efforts to access your biological activities are being carried out.