title: Use of grape pomaces to produce biomass of aKomagataella pastoris strain expressing a bovine chymosin activity.
abstract: The use of agroindustrial wastes not only decreases bioprocesses and disposal costs but also contributes to the upgrading of the residues. An active recombinant methanol-inducible bovine chymosin has been expressed in our laboratory in the yeastKomagataella pastoris, and grape pomace extracts (GRE) were proposed as a convenient C-energy source for the biomass production of the genetically engineered strain. Carbon and nitrogen sources, growth factors, and initial pH conditions were selected by classical methodology; thereafter, growth conditions optimization was performed using statistical designed experiments (DoEs). In the presence of (in g·L(-1)) 67.0 monosaccharides (glucose and fructose) from GRE, 5.0 (NH4)2SO4, and 10.0 sugar cane molasses (CMz), a yield of 20.0 g·L(-1) cell dry weight (CDW) was obtained aerobically after 60 h incubation at 28°C and pH 4.0. Applying a fed-batch strategy with methanol:sorbitol as the enzyme inducers, a chymosin production of 8.53 International Milk Clotting Units (IMCU) per mg protein was obtained in the supernatant. The results presented show that through a statistical design, a simple, cheap, and easy to prepare culture medium could be developed using two agroindustrial derivatives (GRE and CMz) to obtain a higher value added product.
authors.affiliation: Departamento de Ingeniería, Química Facultad de Ingeniería, Universidad de Buenos Aires, Pabellón de Industrias, Ciudad Universitaria Intendente Güiraldes 2160 (1428), Buenos Aires, Argentina.
authors.forename: Diego
authors.initials: D
authors.lastname: Kingston
authors.affiliation: Departamento de Ingeniería, Química Facultad de Ingeniería, Universidad de Buenos Aires, Pabellón de Industrias, Ciudad Universitaria Intendente Güiraldes 2160 (1428), Buenos Aires, Argentina.
authors.forename: Guido F
authors.initials: GF
authors.lastname: Novelli
authors.affiliation: Departamento de Ingeniería, Química Facultad de Ingeniería, Universidad de Buenos Aires, Pabellón de Industrias, Ciudad Universitaria Intendente Güiraldes 2160 (1428), Buenos Aires, Argentina.
authors.forename: Patricia
authors.initials: P
authors.lastname: Cerrutti
authors.affiliation: IIB-INTECH-UNSAM-CONICET 25 de Mayo y Francia, (1650), San Martín, Buenos Aires, Argentina.
authors.forename: Matias N
authors.initials: MN
authors.lastname: Recupero
authors.affiliation: IIB-INTECH-UNSAM-CONICET 25 de Mayo y Francia, (1650), San Martín, Buenos Aires, Argentina.
authors.forename: Martin
authors.initials: M
authors.lastname: Blasco
authors.affiliation: Departamento de Ingeniería, Química Facultad de Ingeniería, Universidad de Buenos Aires, Pabellón de Industrias, Ciudad Universitaria Intendente Güiraldes 2160 (1428), Buenos Aires, Argentina ; IIB-INTECH-UNSAM-CONICET 25 de Mayo y Francia, (1650), San Martín, Buenos Aires, Argentina.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Galvagno
keywords: Agroindustrial by-product; Komagataella pastoris biomass; bovine chymosin