title: Application and bioactive properties of CaTI, a trypsin inhibitor from Capsicum annuum seeds: membrane permeabilization, oxidative stress and intracellular target in phytopathogenic fungi cells.
abstract: BACKGROUND
  During the last few years, a growing number of antimicrobial peptides have been isolated from plants and particularly from seeds. Recent results from our laboratory have shown the purification of a new trypsin inhibitor, named CaTI, from chilli pepper (Capsicum annuum L.) seeds. This study aims to evaluate the antifungal activity and mechanism of action of CaTI on phytopathogenic fungi and detect the presence of protease inhibitors in other species of this genus.
  
  
  RESULTS
  Our results show that CaTI can inhibit the growth of the phytopathogenic fungi Colletotrichum gloeosporioides and C. lindemuthianum. CaTI can also permeabilize the membrane of all tested fungi. When testing the inhibitor on its ability to induce reactive oxygen species, an induction of reactive oxygen species (ROS) and nitric oxide (NO) particularly in Fusarium species was observed. Using CaTI coupled to fluorescein isothiocyanate (FITC), it was possible to determine the presence of the inhibitor inside the hyphae of the Fusarium oxysporum fungus. The search for protease inhibitors in other Capsicum species revealed their presence in all tested species.
  
  
  CONCLUSIONS
  This paper shows the antifungal activity of protease inhibitors such as CaTI against phytopathogenic fungi. Antimicrobial peptides, among which the trypsin protease inhibitor family stands out, are present in different species of the genus Capsicum and are part of the chemical arsenal that plants use to defend themselves against pathogens. © 2017 Society of Chemical Industry.
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Marciele S
authors.initials: MS
authors.lastname: Silva
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Suzanna Ff
authors.initials: SF
authors.lastname: Ribeiro
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Gabriel B
authors.initials: GB
authors.lastname: Taveira
authors.affiliation: Laboratório de Melhoramento e Genética Vegetal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Rosana
authors.initials: R
authors.lastname: Rodrigues
authors.affiliation: Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Katia Vs
authors.initials: KV
authors.lastname: Fernandes
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil.
authors.forename: André O
authors.initials: AO
authors.lastname: Carvalho
authors.affiliation: Laboratório de Toxinas Vegetais, Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, Fortaleza, CE, Brazil.
authors.forename: Ilka Maria
authors.initials: IM
authors.lastname: Vasconcelos
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Erica Oliveira
authors.initials: EO
authors.lastname: Mello
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Valdirene M
authors.initials: VM
authors.lastname: Gomes
mesh_terms: D002212:Capsicum / Q000737:chemistry*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects; D020231:Colletotrichum / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D005659:Fungicides, Industrial / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D005670:Fusarium / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D010935:Plant Diseases / Q000382:microbiology*; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012639:Seeds / Q000737:chemistry*; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D005659:Fungicides, Industrial; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D014361:Trypsin Inhibitors
keywords: Colletotrichum; Fusarium; antimicrobial peptides; protease inhibitor