Recombinant expression, localization and in vitro inhibition of midgut cysteine peptidase (Sl-CathL) from sugarcane weevil, Sphenophorus levis

Cloning and characterization of a basic cysteine-like protease (cathepsin L1) expressed in the gut of larval Diaprepes abbreviatus L. (Coleoptera: Curculionidae)

“…Bown et al (2004) and Fonseca et al (2012) have expressed cathepsin L from the gut of the western corn rootworm (D. virgifera) and the sugarcane weevil, Sphenophorus levis using the methanotropic yeast P. pastoris. The procathepsin L was secreted by the cells into the medium during the fermentation of the cells.…”

“…The procathepsin L was secreted by the cells into the medium during the fermentation of the cells. Bown et al (2004) reported that the secreted proenzyme was cleaved by the yeast proteases and was found as the active cathepsin L, whereas Fonseca et al (2012) reported that the secreted S. levis proenzyme was not activated during the secretion into the medium. Ishidoh and Kominami (1994) expressed procathepsin L in v-Ha-ras transformed NIH3T3 cells and Yamamoto et al (1999) expressed Bombyx mori preprocathepsin L in E. coli.…”

“…The proportion of cathepsin L forms in total cDNA clones is shown as a percentage in parentheses. Fonseca et al, 2012). ClustalW analysis and a phylogenetic tree show that Da-CTSL1 has closer sequence similarity to T. molitor, another coleopteran, than to blood sucking insects like mosquitoes, body louse or lepidopterans (Spodoptera frugiperda and Helicoverpa armigera) (Fig.…”

“…6C). The determination of the pre-pro region of the enzyme was done by homology with other cathepsins (Fonseca et al, 2012). Even though the GST tag kept Da-proCTSL1 soluble in the bacterial cell preventing it from precipitating in the bacterial inclusion bodies, the recombinant enzyme rapidly fell out of solution in low salt LB cultures, during induction and expression at various temperatures (25-37°C), and in the presence of different concentrations of several surfactants (amphipol, CHAPS and OGNG) during bacterial cell lysis.…”

Plant Proteinaceous α-Amylase and Proteinase Inhibitors and Their Use in Insect Pest Control

“…It has been revealed that cathepsin L-like enzymes are the only quantitatively important member of cysteine proteinases presented in midgut of insects. Digestive cathepsin L-like enzymes have been purified from Diabrotica virgifera (Coleoptera: Cucujiformia) (Koiwa et al, 2000), Acyrthosiphon pisum (Hemiptera: Sternorrhyncha) (Cristofoletti et al, 2003), T. molitor (Coleoptera: Cucujiformia) (Cristofoletti et al , 2005), Sphenophorus levis (Coleoptera: Curculionidae) (Soares-Costa et al, 2011;Fonseca et al, 2012), and Triatoma brasiliensis (Reduviidae, Triatominae) (Waniek et al, 2012).…”