Single Step Purification of Bromelain from Ananas Comosus Residues by PEG/Ammonium Sulphate Integrated Aqueous Two-Phase Systems

Diego de Freitas Coelho(1*), Lívia G. Spir(2), Letícia Celia de Lencastre Novaes(3), Priscila G. Mazzola(4), Edgar Silveira(5), Elias B. Tambourgi(6), Adalberto Pessoa Jr.(7)

(1) Faculdade de Engenharia Química, Universidade Estadual de Campinas. Av. Albert Einstein, 500 - CEP 13083-852 - Campinas - SP - Brasil., Brazil
(2) Faculdade de Ciências Médicas, Universidade Estadual de Campinas. Rua Tessália Vieira de Camargo, 126 - CEP 13083-887 - Campinas - SP - Brasil., Brazil
(3) Faculdade de Ciências Farmacêuticas, Universidade de São Paulo. Av. Prof. Lineu Prestes, 580 - CEP 05508-000 -, Brazil
(4) Faculdade de Ciências Médicas, Universidade Estadual de Campinas. Rua Tessália Vieira de Camargo, 126 - CEP 13083-887 - Campinas - SP - Brasil., Brazil
(5) Faculdade de Ciências Farmacêuticas, Universidade de São Paulo. Av. Prof. Lineu Prestes, 580 - CEP 05508-000 -, Brazil
(6) Faculdade de Engenharia Química, Universidade Estadual de Campinas. Av. Albert Einstein, 500 - CEP 13083-852 - Campinas - SP - Brasil., Brazil
(7) Faculdade de Ciências Farmacêuticas, Universidade de São Paulo. Av. Prof. Lineu Prestes, 580 - CEP 05508-000 -, Brazil
(*) Corresponding author


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Abstract


This work evaluates a single step purification of bromelain using a PEG/Ammonium Sulphate integrated aqueous two-phase systems (ATPS). This new process integrates the ammonium sulphate precipitation of unwanted proteins along with purification of target molecule by ATPS. The results, analysed by response surface methodology, showed that higher values of protein yield and enzyme activity recovery could be achieved by desirability optimization. Thus, it was possible to obtain a biological activity recovery over than 87.3% with a purification factor of 11.8 fold, using a system composed by 10.86% PEG 4,000 and 36.21% ammonium sulphate saturation. The purified bromelain showed an increased stability when supplemented with PEG, without any loss of activity after 7 hours of incubation
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Keywords


Bromelain; Ananas Comosus; Aqueous Two-Phase Systems; Purification; Stability

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