title: Lectin I from Bauhinia variegata (BVL-I) expressed by Pichia pastoris inhibits initial adhesion of oral bacteria in vitro.
abstract: Lectins are non-immune proteins that reversibly bind to carbohydrates in a specific manner. Bauhinia variegata lectin I (BVL-I) is a Gal/GalNAc-specific, single-chain lectin isolated from Bauhinia variegata seeds that has been implicated in the inhibition of bacterial adhesion and the healing of damaged skin. Since the source of the native protein (nBVL) is limited, this study aimed to produce recombinant BVL-I in Pichia pastoris (rBVL-Ip). The coding sequence for BVL-I containing preferential codons for P. pastoris was cloned into the pPICZαB plasmid. A single expressing clone was selected and fermented, resulting in the secretion and glycosylation of the protein. Fed-batch fermentation in 7L-scale was performed, and the recombinant lectin was purified from culture supernatant, resulting in a yield of 1.5mg/L culture. Further, rBVL-Ip was compared to nBVL and its recombinant version expressed in Escherichia coli BL21 (DE3) (rBVL-Ie). Although it was expressed as a monomer, rBVL-Ip retained its biological activity since it was able to impair the initial adhesion of Streptococcus mutans and S. sanguinis in an in vitro model of biofilm formation and bacterial adhesion. In summary, rBVL-Ip produced in Pichia pastoris represents a viable alternative to large-scale production, encouraging further biological application studies with this lectin.
authors.affiliation: Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - Campus Capão do Leão, CP 354, CEP 96010-900, Pelotas, RS, Brazil; Faculdade de Medicina/Laboratório de Micologia, Universidade Federal de Rio Grande - Rua General Osório, S/N, CEP 96200-190, Rio Grande, RS, Brazil. Electronic address: gabrielklafke@yahoo.com.br.
authors.forename: Gabriel Baracy
authors.initials: GB
authors.lastname: Klafke
authors.affiliation: Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - Campus Capão do Leão, CP 354, CEP 96010-900, Pelotas, RS, Brazil; Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Spielmannstr. 7, 38106 Braunschweig, Germany. Electronic address: moreira.gmsg@gmail.com.
authors.forename: Gustavo Marçal Schmidt Garcia
authors.initials: GM
authors.lastname: Moreira
authors.affiliation: Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - Campus Capão do Leão, CP 354, CEP 96010-900, Pelotas, RS, Brazil; Faculdade de Medicina/Laboratório de Micologia, Universidade Federal de Rio Grande - Rua General Osório, S/N, CEP 96200-190, Rio Grande, RS, Brazil. Electronic address: julianolacava@gmail.com.
authors.forename: Juliano Lacava
authors.initials: JL
authors.lastname: Pereira
authors.affiliation: Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - Campus Capão do Leão, CP 354, CEP 96010-900, Pelotas, RS, Brazil. Electronic address: bilicadiaz@yahoo.com.br.
authors.forename: Patrícia Diaz
authors.initials: PD
authors.lastname: Oliveira
authors.affiliation: Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - Campus Capão do Leão, CP 354, CEP 96010-900, Pelotas, RS, Brazil. Electronic address: fabriciorc@pop.com.br.
authors.forename: Fabricio Rochedo
authors.initials: FR
authors.lastname: Conceição
authors.affiliation: Faculdade de Odontologia, Universidade Federal de Pelotas - Rua Gonçalves Chaves 457, 96015-560 Pelotas, Rio Grande do Sul, Brazil. Electronic address: rafael.lund@gmail.com.
authors.forename: Rafael Guerra
authors.initials: RG
authors.lastname: Lund
authors.affiliation: Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - Campus Capão do Leão, CP 354, CEP 96010-900, Pelotas, RS, Brazil. Electronic address: grassmann.aa@gmail.com.
authors.forename: André Alex
authors.initials: AA
authors.lastname: Grassmann
authors.affiliation: Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - Campus Capão do Leão, CP 354, CEP 96010-900, Pelotas, RS, Brazil. Electronic address: odirad@terra.com.br.
authors.forename: Odir Antonio
authors.initials: OA
authors.lastname: Dellagostin
authors.affiliation: Núcleo Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - Campus Capão do Leão, CP 354, CEP 96010-900, Pelotas, RS, Brazil. Electronic address: ls_pinto@hotmail.com.
authors.forename: Luciano
authors.initials: L
authors.lastname: da Silva Pinto
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000494:pharmacology*; D001422:Bacterial Adhesion / Q000187:drug effects*; D036481:Bauhinia / Q000737:chemistry*; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli; D006384:Hemagglutination; D006801:Humans; D010843:Pichia / Q000378:metabolism; D037121:Plant Lectins / Q000096:biosynthesis / Q000494:pharmacology*; D011817:Rabbits; D012463:Saliva / Q000382:microbiology; D013295:Streptococcus mutans / Q000187:drug effects; D013298:Streptococcus sanguis / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D037121:Plant Lectins
keywords: Legume lectin; Oral bacteria; Recombinant lectin