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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

Surface plot for biomass production expressed as a function of (NH4)2SO4 and yeast extract (YE) concentrations for the PBSD screening, at level = 0 for the other variables. PBSD, Plackett–Burman screening designs.
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fig02: Surface plot for biomass production expressed as a function of (NH4)2SO4 and yeast extract (YE) concentrations for the PBSD screening, at level = 0 for the other variables. PBSD, Plackett–Burman screening designs.

Mentions: So, the only variables influencing the growth of thisK. pastoris strain positively and significantly were the N and vitamins sources (YE) (Fig.2). On the other hand, the lack of fit of the regression model was not significant, and the parameterFsignificative (<10−3 <0.05) demonstrated a high significance for the regression. At the same time, theF value (19.08) was higher than the critical value (3.48) obtained from tables for a significance level 95% with 5 and 9 degrees of freedom. The quality of fit was determined with the determination coefficientR2 (= 0.914).


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)

Surface plot for biomass production expressed as a function of (NH4)2SO4 and yeast extract (YE) concentrations for the PBSD screening, at level = 0 for the other variables. PBSD, Plackett–Burman screening designs.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig02: Surface plot for biomass production expressed as a function of (NH4)2SO4 and yeast extract (YE) concentrations for the PBSD screening, at level = 0 for the other variables. PBSD, Plackett–Burman screening designs.
Mentions: So, the only variables influencing the growth of thisK. pastoris strain positively and significantly were the N and vitamins sources (YE) (Fig.2). On the other hand, the lack of fit of the regression model was not significant, and the parameterFsignificative (<10−3 <0.05) demonstrated a high significance for the regression. At the same time, theF value (19.08) was higher than the critical value (3.48) obtained from tables for a significance level 95% with 5 and 9 degrees of freedom. The quality of fit was determined with the determination coefficientR2 (= 0.914).

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