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Use of grape pomaces to produce biomass of aKomagataella pastoris strain expressing a bovine chymosin activity.

Kingston D, Novelli GF, Cerrutti P, Recupero MN, Blasco M, Galvagno MA - Food Sci Nutr (2014)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Ingeniería, Química Facultad de Ingeniería, Universidad de Buenos Aires, Pabellón de Industrias, Ciudad Universitaria Intendente Güiraldes 2160 (1428), Buenos Aires, Argentina.

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.

No MeSH data available.


Related in: MedlinePlus

Response surface plot showing the effect of cane molasses (CMz) and (NH4)2SO4 concentrations at 60-h incubation time (+1) on biomass production ofKomagataella pastoris.Inset: predicted versus experimental values for biomass expressed as OD600nm units. The solid line depicted representsy = x.
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fig03: Response surface plot showing the effect of cane molasses (CMz) and (NH4)2SO4 concentrations at 60-h incubation time (+1) on biomass production ofKomagataella pastoris.Inset: predicted versus experimental values for biomass expressed as OD600nm units. The solid line depicted representsy = x.

Mentions: Using the RSM, it was possible to identify the optimum region. Figure3 shows the 3D surface plot described by the regression model taking into account the influence of CMz (replacing YE as a source of vitamins according to Fig.2) and (NH4)2SO4 concentrations, at a constant coded level +1(=60 h) for incubation time. This plot shows that in the presence of maximal GRE concentrations, optimal biomass production could be obtained at the highest levels (+1) of both (NH4)2SO4 and CMz. Higher values of both CMz and (NH4)2SO4 were assayed, but no significant increase in biomass production was attained.


Use of grape pomaces to produce biomass of aKomagataella pastoris strain expressing a bovine chymosin activity.

Kingston D, Novelli GF, Cerrutti P, Recupero MN, Blasco M, Galvagno MA - Food Sci Nutr (2014)

Response surface plot showing the effect of cane molasses (CMz) and (NH4)2SO4 concentrations at 60-h incubation time (+1) on biomass production ofKomagataella pastoris.Inset: predicted versus experimental values for biomass expressed as OD600nm units. The solid line depicted representsy = x.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256579&req=5

fig03: Response surface plot showing the effect of cane molasses (CMz) and (NH4)2SO4 concentrations at 60-h incubation time (+1) on biomass production ofKomagataella pastoris.Inset: predicted versus experimental values for biomass expressed as OD600nm units. The solid line depicted representsy = x.
Mentions: Using the RSM, it was possible to identify the optimum region. Figure3 shows the 3D surface plot described by the regression model taking into account the influence of CMz (replacing YE as a source of vitamins according to Fig.2) and (NH4)2SO4 concentrations, at a constant coded level +1(=60 h) for incubation time. This plot shows that in the presence of maximal GRE concentrations, optimal biomass production could be obtained at the highest levels (+1) of both (NH4)2SO4 and CMz. Higher values of both CMz and (NH4)2SO4 were assayed, but no significant increase in biomass production was attained.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Ingeniería, Química Facultad de Ingeniería, Universidad de Buenos Aires, Pabellón de Industrias, Ciudad Universitaria Intendente Güiraldes 2160 (1428), Buenos Aires, Argentina.

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.

No MeSH data available.


Related in: MedlinePlus