Aqueous biphasic systems (ABSs) are an interesting alternative for separating industrial enzymes due to easy scale-up and low operational cost. The proteases of Pseudomonas sp. M211 were purified through ABS platforms formed by polyethylene glycol (PEG) and citrate buffer salt. Two experimental designs 23 + 4 were performed to evaluate the following parameters: molar mass of PEG (MPEG), concentration of PEG (CPEG), concentration of citrate buffer (CCit), and pH. The partition coefficient (K), activity yield (Y), and purification factor (PF) were the responses analyzed. The best purification performance was obtained with the system composed of MPEG = 10,000 g/mol, CPEG = 22 wt%, CCit = 12 wt%, pH = 8.0; the responses obtained were K = 4.9, Y = 84.5%, PF = 15.1, and tie-line length = 52.74%. The purified proteases of Pseudomonas sp. (PPP) were used to obtain hydrolysates of Lupinus mutabilis (Peruvian lupin cultivar) seed protein in comparison with the commercial protease Alcalase® 2.4L. A strong correlation between hydrolysis degree and radical scavenging activity was observed, and the highest antioxidant activity was obtained with Alcalase® (1.40 and 3.47 μmol Trolox equivalent/mg protein, for 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) and oxygen radical absorbance capacity, respectively) compared with PPP (0.55 and 1.03 μmol Trolox/mg protein). Nevertheless, the IC50 values were lower than those often observed for antioxidant hydrolysates from plant proteins. PEG/citrate buffer system is valuable to purify Pseudomonas proteases from the fermented broth, and the purified protease could be promising to produce antioxidant protein hydrolysates.
Bibliographical noteFunding Information:
This work was supported by “Consejo Nacional de Ciencia, Tecnología e Innovación Tecnológica” (CONCYTEC), Peru (Financial Agreement Number 007‐2014‐FONDECYT). A. Brandelli is research awardee of Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brazil) (grant 306936/2017‐8). J. H. P. M. S. thank the financial support given by FAPESP within the postdoctoral grant 2018/25994‐2.
Consejo Nacional de Ciencia y Tecnología, Grant/Award Number: 007‐2014‐FONDECYT; Conselho Nacional de Desenvolvimento Científico e Tecnológico, Grant/Award Number: 306936/2017‐8; Fundação de Amparo à Pesquisa do Estado de São Paulo, Grant/Award Number: 2018/25994‐2; Fundação para a Ciência e a Tecnologia, Grant/Award Number: SFRH/BD/102915/2014 Funding information
This work was supported by ?Consejo Nacional de Ciencia, Tecnolog?a e Innovaci?n Tecnol?gica? (CONCYTEC), Peru (Financial Agreement Number 007-2014-FONDECYT). A. Brandelli is research awardee of Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (Brazil) (grant 306936/2017-8). J.?H.?P.?M.?S. thank the financial support given by FAPESP within the postdoctoral grant 2018/25994-2.
© 2020 American Institute of Chemical Engineers
- Lupinus mutabilis
- Pseudomonas sp.
- aqueous biphasic systems