Lectin I from Bauhinia variegata (BVL-I) expressed by Pichia pastoris inhibits initial adhesion of oral bacteria in vitro

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• 作者：Gabriel Baracy Klafke^a ; ^b ; ^{gabrielklafke@yahoo.com.br" class="auth_mail" title="E-mail the corresponding author} ; Gustavo Març ; al Schmidt Garcia Moreira^a ; ^c ; ^{moreira.gmsg@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; Juliano Lacava Pereira^a ; ^b ; ^{julianolacava@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; Patrí ; cia Diaz Oliveira^a ; ^{bilicadiaz@yahoo.com.br" class="auth_mail" title="E-mail the corresponding author} ; Fabricio Rochedo Conceiç ; ã ; o^a ; ^{fabriciorc@pop.com.br" class="auth_mail" title="E-mail the corresponding author} ; Rafael Guerra Lund^d ; ^{rafael.lund@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; André ; Alex Grassmann^a ; ^{grassmann.aa@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; Odir Antonio Dellagostin^a ; ^{odirad@terra.com.br" class="auth_mail" title="E-mail the corresponding author} ; Luciano da Silva Pinto^a ; ^{ls_pinto@hotmail.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Legume lectin ; Recombinant lectin ; Oral bacteria
• 刊名：International Journal of Biological Macromolecules
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：93
• 期：part_PA
• 页码：913-918
• 全文大小：1108 K

文摘

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 7 L-scale was performed, and the recombinant lectin was purified from culture supernatant, resulting in a yield of 1.5 mg/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.