Examinando por Autor "Pillaca-Pullo, Omar"
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Publicación Acceso abierto “Effects of Oxygen Transference on Protease Production by Rhodotorula mucilaginosa CBMAI 1528 in a Stirred Tank Bioreactor “(MDPI, 2022-11-15) Machado, Suellen; Feitosa, Valker; Pillaca-Pullo, Omar; Lario, Luciana; Sette, Lara; Pessoa, Jr., Adalberto; Alves, Harley“Microbial proteases, especially aspartic proteases, are an essential group of enzymes produced from different microorganisms. Microbial proteases have several applications, mainly in the food, beverage, cosmetic, and pharmaceutical industries, due to their efficiency in the processing and in the manufacturing stages. The yeast Rhodotorula mucilaginosa CBMAI 1528 was isolated from the Antarctic environment and was previously reported to have higher extracellular aspartic protease production. In addition, advances in the operational conditions of bioreactors for enzyme production are important to reduce the gap associated with scaling−up processes. This is the first study that evaluates the influence of oxygen transference (kLa) on the protease production of R. mucilaginosa yeast. To that end, batch cultures were created in a stirred tank bioreactor using Sabouraud dextrose broth at 25 ◦C for 72 h under kLa values from 18 to 135 h−1 . The results show that kLa (121 h−1 ) obtained at 500 rpm and 1.5 vvm plays an important role in protease production (124.9 U/mL) and productivity (6.784 U/L.h) as well as biomass (10.4 g/L), µmax (0.14 h−1 ) and Yx/s (0.484 g/g). In conclusion, R. mucilaginosa showed high yield production in aerobic culture with the efficiency of protease expression and secretion influenced by kLa. In this sense, our results could be used for further industrial investment.“Publicación Acceso abierto “Endochitinase and Chitobiosidase Production by Marine Aeromonas caviae CHZ306: Establishment of Nitrogen Supplementation“(MDPI, 2023-03-09) Cardozo, Flavio; Feitosa, Valker; Pillaca-Pullo, Omar; Pessoa, Adalberto“Aeromonas caviae CHZ306, a marine-derived bacterium isolated from zooplankton, can use chitin (a polymer of a β-(1,4)-linked N-acetyl-D-glucosamine) as a carbon source. The chitin is hydrolyzed by chitinolytic enzymes, namely endochitinases and exochitinases (chitobiosidase and N-acetyl-glucosaminidase). Indeed, the chitinolytic pathway is initiated by the coexpression of the enzymes endochitinase (EnCh) and chitobiosidase (ChB); however, few studies, including biotechnological production of these enzymes, have been reported, although chitosaccharide are helpful in several industries, such as cosmetics. This study demonstrates the potential to maximize the simultaneous EnCh and ChB production by nitrogen supplementation on culture media. Twelve different nitrogen supplementation sources (inorganic and organic) previously analyzed in elemental composition (carbon and nitrogen) were tested and evaluated in the Erlenmeyer flask culture of A. caviae CHZ306 for EnCh and ChB expression. None of the nutrients inhibited bacterial growth, and the maximum activity in both EnCh and ChB was observed at 12 h, using corn-steep solids and peptone A. Corn-steep solids and peptone A were then combined at three ratios (1:1, 1:2, and 2:1) to maximize the production. The high activities for EnCh (30.1 U.L−1 ) and ChB (21.3 U.L−1 ) were obtained with 2:1 corn-steep solids and peptone A, corresponding to more than 5- and 3-fold enhancement, respectively, compared to the control condition.“Publicación Acceso abierto “Endochitinase and Chitobiosidase Production by Marine Aeromonas caviae CHZ306: Establishment of Nitrogen Supplementation“(MDPI, 2023-03-29) Cardozo, Flavio; Feitosa, Valker; Pillaca-Pullo, Omar; Pessoa, Adalberto“Aeromonas caviae CHZ306, a marine-derived bacterium isolated from zooplankton, can use chitin (a polymer of a β-(1,4)-linked N-acetyl-D-glucosamine) as a carbon source. The chitin is hydrolyzed by chitinolytic enzymes, namely endochitinases and exochitinases (chitobiosidase and N-acetyl-glucosaminidase). Indeed, the chitinolytic pathway is initiated by the coexpression of the enzymes endochitinase (EnCh) and chitobiosidase (ChB); however, few studies, including biotechnological production of these enzymes, have been reported, although chitosaccharide are helpful in several industries, such as cosmetics. This study demonstrates the potential to maximize the simultaneous EnCh and ChB production by nitrogen supplementation on culture media. Twelve different nitrogen supplementation sources (inorganic and organic) previously analyzed in elemental composition (carbon and nitrogen) were tested and evaluated in the Erlenmeyer flask culture of A. caviae CHZ306 for EnCh and ChB expression. None of the nutrients inhibited bacterial growth, and the maximum activity in both EnCh and ChB was observed at 12 h, using corn-steep solids and peptone A. Corn-steep solids and peptone A were then combined at three ratios (1:1, 1:2, and 2:1) to maximize the production. The high activities for EnCh (30.1 U.L−1 ) and ChB (21.3 U.L−1 ) were obtained with 2:1 corn-steep solids and peptone A, corresponding to more than 5- and 3-fold enhancement, respectively, compared to the control condition.“Publicación Acceso abierto Scale-Up of Capsular Polysaccharide Production Process by Haemophilus influenzae Type b Using kLa Criterion(MDPI, 2022-08-25) Pillaca-Pullo, Omar; Dias Vieira , Lucas; Takagi , MickiePolyribosyl-ribitol-phosphate (PRP) from Haemophilus influenzae type b (Hib) is an active immunizing molecule used in the production of the vaccine against H. influenzae, and industrial production could contribute to satisfying a world demand especially in developing countries. In this sense, the aim of this study was to establish a scale-up process using the constant oxygen mass transfer coefficient (kLa) such as the criterion for production of PRP in three different sizes of bioreactor systems. Three different kLa values (24, 52 and 80 h−1) were evaluated in which the biological influence in a 1.5 L bioreactor and 52 h−1 was selected to scale-up the production process until a 75 L pilot-scale bioreactor was achieved. Finally, the fed-batch phase was started under a dissolved oxygen concentration (pO2) at 30% of the saturation in the 75 L bioreactor to avoid oxygen limitation; the performance of production presented high efficiency (9.0 g/L DCW-dry cell weight and 1.4 g/L PRP) in comparison with previous scale-up studies. The yields, productivity and kinetic behavior were similar in the three-size bioreactor systems in the batch mode indicating that kLa is possible to use for PRP production at large scales. This process operated under two stages and successfully produced DCW and PRP in the pilot scale and could be beneficial for future bioprocess operations that may lead to higher production and less operative cost. View Full-TextPublicación Acceso abierto Scale-Up of Capsular Polysaccharide Production Process by Haemophilus influenzae Type b Using kLa Criterion(MDPI, 2022-08-25) Pillaca-Pullo, Omar; Dias Vieira, Lucas; Takagi, MickiePolyribosyl-ribitol-phosphate (PRP) from Haemophilus influenzae type b (Hib) is an active immunizing molecule used in the production of the vaccine against H. influenzae, and industrial production could contribute to satisfying a world demand especially in developing countries. In this sense, the aim of this study was to establish a scale-up process using the constant oxygen mass transfer coefficient (kLa) such as the criterion for production of PRP in three different sizes of bioreactor systems. Three different kLa values (24, 52 and 80 h−1) were evaluated in which the biological influence in a 1.5 L bioreactor and 52 h−1 was selected to scale-up the production process until a 75 L pilot-scale bioreactor was achieved. Finally, the fed-batch phase was started under a dissolved oxygen concentration (pO2) at 30% of the saturation in the 75 L bioreactor to avoid oxygen limitation; the performance of production presented high efficiency (9.0 g/L DCW-dry cell weight and 1.4 g/L PRP) in comparison with previous scale-up studies. The yields, productivity and kinetic behavior were similar in the three-size bioreactor systems in the batch mode indicating that kLa is possible to use for PRP production at large scales. This process operated under two stages and successfully produced DCW and PRP in the pilot scale and could be beneficial for future bioprocess operations that may lead to higher production and less operative cost.
