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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

Effect of growth ofKomagataella pastoris in GRE (A) with different N source:  GRE without supplementation;  GRE + meat peptone,  GRE + tryptone;  GRE + (NH4)2SO4;  GRE + corn steep liquor;  GRE + urea; (B) growth factors addition:  GRE without supplements;  GRE + yeast extract;  GRE + cane molasses;  GRE + biotin; (C) at different pH values.  pH = 3,  pH = 4,  pH = 5,  pH = 6. GRE, grape pomace extracts.
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fig01: Effect of growth ofKomagataella pastoris in GRE (A) with different N source: GRE without supplementation; GRE + meat peptone, GRE + tryptone; GRE + (NH4)2SO4; GRE + corn steep liquor; GRE + urea; (B) growth factors addition: GRE without supplements; GRE + yeast extract; GRE + cane molasses; GRE + biotin; (C) at different pH values. pH = 3, pH = 4, pH = 5, pH = 6. GRE, grape pomace extracts.

Mentions: As Figure1A shows, yeast growth was enhanced when N (organic or inorganic) substrates were added. Because addition of the N sources used increased both maximal-specific growth rate and biomass concentration reached in the stationary phase of growth, GRE was considered deficient in N for Clone 1 growth. Curves depicted in Figure1 showed that the inorganic salt (NH4)2SO4 was as effective as meat peptone, tryptone, and CSL ones with respect to the highest biomass productions and productivities obtained at 48 h incubation.


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)

Effect of growth ofKomagataella pastoris in GRE (A) with different N source:  GRE without supplementation;  GRE + meat peptone,  GRE + tryptone;  GRE + (NH4)2SO4;  GRE + corn steep liquor;  GRE + urea; (B) growth factors addition:  GRE without supplements;  GRE + yeast extract;  GRE + cane molasses;  GRE + biotin; (C) at different pH values.  pH = 3,  pH = 4,  pH = 5,  pH = 6. GRE, grape pomace extracts.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig01: Effect of growth ofKomagataella pastoris in GRE (A) with different N source: GRE without supplementation; GRE + meat peptone, GRE + tryptone; GRE + (NH4)2SO4; GRE + corn steep liquor; GRE + urea; (B) growth factors addition: GRE without supplements; GRE + yeast extract; GRE + cane molasses; GRE + biotin; (C) at different pH values. pH = 3, pH = 4, pH = 5, pH = 6. GRE, grape pomace extracts.
Mentions: As Figure1A shows, yeast growth was enhanced when N (organic or inorganic) substrates were added. Because addition of the N sources used increased both maximal-specific growth rate and biomass concentration reached in the stationary phase of growth, GRE was considered deficient in N for Clone 1 growth. Curves depicted in Figure1 showed that the inorganic salt (NH4)2SO4 was as effective as meat peptone, tryptone, and CSL ones with respect to the highest biomass productions and productivities obtained at 48 h incubation.

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