title: Production of orotic acid by a Klura3Δ mutant of Kluyveromyces lactis.
abstract: We demonstrated that a Klura3Δ, mutant of the yeast Kluyveromyces lactis is able to produce and secrete into the growth medium considerable amounts of orotic acid. Using yeast extract-peptone-glucose (YPD) based media we optimized production conditions in flask and bioreactor cultures. With cells grown in YPD 5% glucose medium, the best production in flask was obtained with a 1:12.5 ratio for flask: culture volume, 180 rpm, 28°C and 200 mM MOPS for pH stabilization at neutral values (initial culture pH at 8.0). The best production in a 2 L bioreactor was achieved at 500 rpm with 1 vvm aeration, 28°C and pH 7.0. Under these optimum conditions, similar rates of orotic acid production were obtained and maximum concentration achieved after 96 h was 6.7 g/L in flask and bioreactor cultures. These results revealed an excellent reproducibility between both systems and provided evidence for the biotechnological potential of Klura3Δ strain to produce orotic acid since the amounts obtained are comparable to the production in flask using a similar mutant of the industrially valuable Corynebacterium glutamicum.
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen s/n, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Rua do Campo Alegre 823, 4150-180 Porto, Portugal; Instituto de Ciências Biomédicas Abel Salazar, Departamento de Biologia Molecular, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
authors.forename: Nuno
authors.initials: N
authors.lastname: Carvalho
authors.affiliation: Centro de Engenharia Biológica, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Coelho
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen s/n, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Rua do Campo Alegre 823, 4150-180 Porto, Portugal; Instituto de Ciências Biomédicas Abel Salazar, Departamento de Biologia Molecular, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
authors.forename: Luís
authors.initials: L
authors.lastname: Gales
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen s/n, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Rua do Campo Alegre 823, 4150-180 Porto, Portugal; Instituto de Ciências Biomédicas Abel Salazar, Departamento de Biologia Molecular, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
authors.forename: Vítor
authors.initials: V
authors.lastname: Costa
authors.affiliation: Centro de Engenharia Biológica, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal.
authors.forename: José António
authors.initials: JA
authors.lastname: Teixeira
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen s/n, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Rua do Campo Alegre 823, 4150-180 Porto, Portugal; Instituto de Ciências Biomédicas Abel Salazar, Departamento de Biologia Molecular, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal. Electronic address: pmferrei@ibmc.up.pt.
authors.forename: Pedro
authors.initials: P
authors.lastname: Moradas-Ferreira
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D019149:Bioreactors / Q000382:microbiology*; D001709:Biotechnology; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007716:Kluyveromyces / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D060847:Metabolic Engineering; D009154:Mutation; D009963:Orotic Acid / Q000378:metabolism*; D015203:Reproducibility of Results; D013696:Temperature; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D009963:Orotic Acid
keywords: Bioprocess optimization; KlURA3; Kluyveromyces lactis; Orotic acid; Yeast biotechnology