title: Generation of Lactococcus lactis capable of coexpressing epidermal growth factor and trefoil factor to enhance in vitro wound healing.
abstract: Epidermal growth factor (EGF) and trefoil factor 3 (TFF3) are peptides that actively support the restitution and repair of mucosal epithelial barriers. Previous studies have shown that TFF3 enhanced EGF effect in wound healing, suggesting that the combined application of the two factors may be advantageous in clinical tissue repair. Expression of multiple proteins in a single host is a desirable approach in a biotechnological process, allowing to reduce cost and increase production efficiency. The aim of the present study was to study the feasibility of coexpressing EGF and TFF3 in food grade bacteria, Lactococcus lactis (L. lactis). Using an expression construct allowing simultaneous translation of two separate recombinant peptides, we generated a L. lactis that coexpressed and secreted EGF and TFF3 dually (LL-ET). Western blot analysis revealed that LL-ET secreted 45-54 % more total recombinant peptides (EGF+TFF3) per flask fermentation and 21-37 % more total recombinant proteins in bioreactor fermentation compared to their single factor expressing L. lactis counterparts (LL-EGF and LL-TFF3, respectively). The resulted recombinant EGF and TFF3 showed enhancement in wound healing activity in vitro. Our data suggest that the dual expression and secretion of EGF and TFF3 by L. lactis effectively accelerated cell migration, demonstrating potential future oral application of L. lactis fermentation product containing dual factors or a cocktail of factors to potentially treat intestinal damage and inflammation.
authors.affiliation: Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
authors.forename: Evanna
authors.initials: E
authors.lastname: Huynh
authors.forename: Julang
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D001688:Biological Products / Q000378:metabolism*; D019149:Bioreactors; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D015870:Gene Expression; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013552:Swine; D000071165:Trefoil Factor-3
chemical_list: D001688:Biological Products; D010455:Peptides; D011994:Recombinant Proteins; C082495:TFF3 protein, human; D000071165:Trefoil Factor-3; D004815:Epidermal Growth Factor