title: BbrzSP-32, the first serine protease isolated from Bothrops brazili venom: Purification and characterization.
abstract: Snake venom toxins are related not only in detention, death and the promotion of initial digestion of prey but also due to their different biochemical, structural and pharmacological effects they can result in new drugs. Among these toxins snake venom serine proteases (SVSPs) should be highlighted because they are responsible for inducing changes in physiological functions such as blood coagulation, fibrinolysis, and platelet aggregation. This article presents the first serine protease (SP) isolated from Bothrops brazili: BbrzSP-32. The new SP showed 36 kDa of relative molecular mass and its absolute mass was confirmed by mass spectrometry as 32,520 Da. It presents 79.48% identity when compared to other SVSPs and was able to degrade the α-chain of fibrinogen, in in vitro models, because of this it is considered a SVTLE-A. It showed dose-dependent activity in the process of degradation of fibrin networks demonstrating greater specificity for this activity when compared to its thrombolytic action. BbrzSP-32 demonstrated proteolytic activity on gelatin and chromogenic substrates for serine proteases and thrombin-like enzymes (S-2288 and S-2238 respectively), besides having coagulant activity on human plasma. After pre-incubation with PMSF and benzamidine the coagulant and proteolytic activities on the S-2288 and S-2238 substrates were reduced. BbrzSP-32 shows stability against pH and temperature variations, demonstrating optimum activity between 30 and 40 °C and in the pH range 7.5 to 8.5. A new SP with potential biotechnological application was isolated.
authors.affiliation: Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil.
authors.forename: Kayena D
authors.initials: KD
authors.lastname: Zaqueo
authors.affiliation: Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil.
authors.forename: Anderson M
authors.initials: AM
authors.lastname: Kayano
authors.affiliation: Universidade Federal Fluminense, UFF, Niterói, RJ, Brazil.
authors.forename: Thaisa F S
authors.initials: TF
authors.lastname: Domingos
authors.affiliation: Universidade Federal Fluminense, UFF, Niterói, RJ, Brazil.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Moura
authors.affiliation: Universidade Federal Fluminense, UFF, Niterói, RJ, Brazil.
authors.forename: André L
authors.initials: AL
authors.lastname: Fuly
authors.affiliation: Universidade Regional Amazónica IKIAM, Via Muyuna Km 7, Tena, Napo, Ecuador.
authors.forename: Saulo L
authors.initials: SL
authors.lastname: da Silva
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), 08028-Barcelona, Spain and CIBER-BBN, Barcelona Science Park, Barcelona 08028, Spain.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Acosta
authors.affiliation: Proteomic Platform, Barcelona Science Park, Barcelona 08028, Spain.
authors.forename: Eliandre
authors.initials: E
authors.lastname: Oliveira
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), 08028-Barcelona, Spain and CIBER-BBN, Barcelona Science Park, Barcelona 08028, Spain; Department of Organic Chemistry University of Barcelona, Barcelona 08028, Spain; School of Chemistry, University of KwaZulu Natal, Durban 4001, South Africa.
authors.forename: Fernando
authors.initials: F
authors.lastname: Albericio
authors.affiliation: Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil.
authors.forename: Fernando B
authors.initials: FB
authors.lastname: Zanchi
authors.affiliation: Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil.
authors.forename: Juliana P
authors.initials: JP
authors.lastname: Zuliani
authors.affiliation: Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil.
authors.forename: Leonardo A
authors.initials: LA
authors.lastname: Calderon
authors.affiliation: Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil. Electronic address: stabeli@fiocruz.br.
authors.forename: Rodrigo G
authors.initials: RG
authors.lastname: Stábeli
authors.affiliation: Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Fundação Oswaldo Cruz, FIOCRUZ Rondônia e Departamento de Medicina, Universidade Federal de Rondônia, Brazil. Electronic address: andreimar@fiocruz.br.
authors.forename: Andreimar M
authors.initials: AM
authors.lastname: Soares
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops; D003435:Crotalid Venoms / Q000737:chemistry*; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D057057:Serine Proteases / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D003435:Crotalid Venoms; D057057:Serine Proteases
keywords: Biotechnology; Enzyme; Snake venom; Thrombin-like; Viperidae