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Identification of four novel members of Kunitz-like α-amylase inhibitors family from Delonix regia with activity toward Coleopteran insects

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Title: Identification of four novel members of Kunitz-like α-amylase inhibitors family from Delonix regia with activity toward Coleopteran insects
Authors: Alves, Debora T.1, Vasconcelos, Ilka M.2, Oliveira, José T.A.2, Farias, Luciana R.1, Dias, Simoni C.1, Chiarello, Marileusa D.1, Maria-Neto, Simone1, Franco, Octávio L.1,3 ocfranco@pos.ucb.br
Source: Pesticide Biochemistry & Physiology. Nov2009, Vol. 95 Issue 3, p166-172. 7p.
Subject Terms: *Crop improvement, *Plant defenses, Amylase inhibitors, Royal poinciana, Enzyme inhibitors, Polyacrylamide gel electrophoresis, High performance liquid chromatography
Abstract: Abstract: Crop improvement generally focuses on yield, seed quality and nutritional characteristics, rather than resistance to biotic and abiotic stresses. A clear consequence of this approach is the absence of natural anti-feedant toxins in some improved seed materials, allowing predation of commercial crops by insect herbivores. Cowpea (Vigna unguiculata), commonly cultivated by small farmers, is particularly affected by insect-pests that reproduce and develop inside stored seeds. One alternative to conventional pesticides for pest control is the use of biotechnological tools, such as the digestive enzyme inhibitors, that could be introduced in transgenic crops to enhance resistance. In this study, it was verified that the in vivo bioassays using artificial seeds containing 0.5%, 1.0% and 1.5% (w/w) of Delonix regia rich fraction, containing α-amylase inhibitors with effectiveness toward insect α-amylases and other sources, caused remarkable reduction in development and increased mortality of Callosobruchus maculatus cowpea weevil and to cotton boll weevil Anthonomus grandis. Therefore, attempts were made to isolated those inhibitors by SP-Sepharose ion exchange chromatography followed by high performance liquid chromatography on a Vydac C18-TP analytical column. Four inhibitor peaks were obtained with molecular masses of 6.0, 20 and 24kDa. Their N-termini showed high sequence similarities with Kunitz-like inhibitor family members. These results provide evidence that D. regia synthesizes a multiple family of Kunitz-like α-amylase inhibitors, with different molecular masses and a wide biotechnological potential to control insect-pests. [Copyright &y& Elsevier]
Copyright of Pesticide Biochemistry & Physiology is the property of Academic Press Inc. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Identification of four novel members of Kunitz-like α-amylase inhibitors family from Delonix regia with activity toward Coleopteran insects
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  Data: <searchLink fieldCode="AR" term="%22Alves%2C+Debora+T%2E%22">Alves, Debora T.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Vasconcelos%2C+Ilka+M%2E%22">Vasconcelos, Ilka M.</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Oliveira%2C+José+T%2EA%2E%22">Oliveira, José T.A.</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Farias%2C+Luciana+R%2E%22">Farias, Luciana R.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Dias%2C+Simoni+C%2E%22">Dias, Simoni C.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Chiarello%2C+Marileusa+D%2E%22">Chiarello, Marileusa D.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Maria-Neto%2C+Simone%22">Maria-Neto, Simone</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Franco%2C+Octávio+L%2E%22">Franco, Octávio L.</searchLink><relatesTo>1,3</relatesTo><i> ocfranco@pos.ucb.br</i>
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  Data: <searchLink fieldCode="JN" term="%22Pesticide+Biochemistry+%26+Physiology%22">Pesticide Biochemistry & Physiology</searchLink>. Nov2009, Vol. 95 Issue 3, p166-172. 7p.
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  Data: *<searchLink fieldCode="DE" term="%22Crop+improvement%22">Crop improvement</searchLink><br />*<searchLink fieldCode="DE" term="%22Plant+defenses%22">Plant defenses</searchLink><br /><searchLink fieldCode="DE" term="%22Amylase+inhibitors%22">Amylase inhibitors</searchLink><br /><searchLink fieldCode="DE" term="%22Royal+poinciana%22">Royal poinciana</searchLink><br /><searchLink fieldCode="DE" term="%22Enzyme+inhibitors%22">Enzyme inhibitors</searchLink><br /><searchLink fieldCode="DE" term="%22Polyacrylamide+gel+electrophoresis%22">Polyacrylamide gel electrophoresis</searchLink><br /><searchLink fieldCode="DE" term="%22High+performance+liquid+chromatography%22">High performance liquid chromatography</searchLink>
– Name: Abstract
  Label: Abstract
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  Data: Abstract: Crop improvement generally focuses on yield, seed quality and nutritional characteristics, rather than resistance to biotic and abiotic stresses. A clear consequence of this approach is the absence of natural anti-feedant toxins in some improved seed materials, allowing predation of commercial crops by insect herbivores. Cowpea (Vigna unguiculata), commonly cultivated by small farmers, is particularly affected by insect-pests that reproduce and develop inside stored seeds. One alternative to conventional pesticides for pest control is the use of biotechnological tools, such as the digestive enzyme inhibitors, that could be introduced in transgenic crops to enhance resistance. In this study, it was verified that the in vivo bioassays using artificial seeds containing 0.5%, 1.0% and 1.5% (w/w) of Delonix regia rich fraction, containing α-amylase inhibitors with effectiveness toward insect α-amylases and other sources, caused remarkable reduction in development and increased mortality of Callosobruchus maculatus cowpea weevil and to cotton boll weevil Anthonomus grandis. Therefore, attempts were made to isolated those inhibitors by SP-Sepharose ion exchange chromatography followed by high performance liquid chromatography on a Vydac C18-TP analytical column. Four inhibitor peaks were obtained with molecular masses of 6.0, 20 and 24kDa. Their N-termini showed high sequence similarities with Kunitz-like inhibitor family members. These results provide evidence that D. regia synthesizes a multiple family of Kunitz-like α-amylase inhibitors, with different molecular masses and a wide biotechnological potential to control insect-pests. [Copyright &y& Elsevier]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Pesticide Biochemistry & Physiology is the property of Academic Press Inc. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1016/j.pestbp.2009.08.011
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      – SubjectFull: High performance liquid chromatography
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              Text: Nov2009
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