02785nas a2200265 4500008004100000022001400041245014000055210006900195260007400264300001200338490000700350520181600357653001302173653002002186653001502206653002502221653001102246653002502257100001902282700002002301700002502321700002202346700002102368856013002389 2005 eng d a0167-441200aKunitz trypsin inhibitor from chickpea (Cicer arietinum L.) that exerts anti-metabolic effect on podborer (Helicoverpa armigera) larvae0 aKunitz trypsin inhibitor from chickpea Cicer arietinum L that ex aVAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDSbSPRINGERcFEB a359-3740 v573 a
Chickpea (Cicer arietinum L.) seeds contain Bowman-Birk proteinase inhibitors, which are ineffective against the digestive proteinases of larvae of the insect pest Helicoverpa armigera. We have identified and purified a low expressing proteinase inhibitor (PI), distinct from the Bowman-Birk Inhibitors and active against H. armigera gut proteinases (HGP), from chickpea seeds. N-terminal sequencing of this HGP inhibitor revealed a sequence similar to reported pea (Pisum sativum) and chickpea alpha-L-fucosidases and also homologous to legume Kunitz inhibitors. The identity was confirmed by matrix assisted laser desorption ionization - time of flight analysis of tryptic peptides and isolation of DNA sequence coding for the mature protein. Available sequence data showed that this protein forms a distinct phylogenetic cluster with Kunitz inhibitors from Glycine max, Medicago truncatula, P. sativum and Canavalia lineata. The isolated coding sequence was cloned into a yeast expression vector and produced as a recombinant protein in Pichia pastoris. alpha-L-fucosidase activity was not detectable in purified or recombinant protein, by solution assays. The recombinant protein did not inhibit chymotrypsin or subtilisin activity but did exhibit stoichiometric inhibition of trypsin, comparable to soybean Kunitz trypsin inhibitor. The recombinant protein exhibited higher inhibition of total HGP activity as compared to soybean kunitz inhibitor, even though it preferentially inhibited HGP-trypsins. H. armigera larvae fed on inhibitor-incorporated artificial diet showed significant reduction in average larval weight after 18 days of feeding demonstrating potent antimetabolic activity. The over-expression of this gene in chickpea could act as an endogenous source of resistance to H. armigera.
10aChickpea10aCicer arietinum10afucosidase10aHelicoverpa armigera10aKunitz10aProteinase inhibitor1 aSrinivasan, A.1 aGiri, Ashok, P.1 aHarsulkar, Abhay, M.1 aGatehouse, J., A.1 aGupta, Vidya, S. uhttp://library.ncl.res.in/content/kunitz-trypsin-inhibitor-chickpea-cicer-arietinum-l-exerts-anti-metabolic-effect-podborer-0