Partition and recovery of phytase from Absidia blakesleeana URM5604 using PEG–citrate aqueous two-phase systems

► Phytase was partition and recovery from crude extract in a single-step operation. ► For this, ATPS composed by PEG–citrate was utilized. ► To optimize the conditions was used 24 factorial design. ► It was possible to obtain a partition (14.42) with recovery (115.24%). ► ATPS is an interesting and...

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Bibliographic Details
Published in:Fluid phase equilibria 2012-03, Vol.318, p.34-39
Main Authors: Neves, Maria Luiza Carvalho, Porto, Tatiana Souza, Souza-Motta, Cristina Maria, Spier, Michele Rigon, Soccol, Carlos Ricardo, Moreira, Keila Aparecida, Porto, Ana Lúcia Figueiredo
Format: Article
Language:English
Subjects:
Aqueous two-phase system
Chemistry
Exact sciences and technology
General and physical chemistry
Partition
PEG/citrate
Phase equilibria
Phytase
Recovery
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Summary:► Phytase was partition and recovery from crude extract in a single-step operation. ► For this, ATPS composed by PEG–citrate was utilized. ► To optimize the conditions was used 24 factorial design. ► It was possible to obtain a partition (14.42) with recovery (115.24%). ► ATPS is an interesting and efficient alternative to partition and recovery of phytase. The partition and recovery behavior of phytase, produced by solid-state cultivation utilizing citrus pulp as substrate, was determined in an aqueous two-phase system (ATPS), composed by polyethylene glycol (PEG–citrate). The 24 factorial design was conducted to evaluate the effect of the following independent variables: PEG molar mass, PEG and citrate concentrations and pH, while partition coefficient, recovery and purification factor were selected as the variables responses. The best recovery in phytase activity (115.24%) condition was obtained with 26% (w/w) PEG 8000 (g/mol), 12% (w/w) sodium citrate at pH 8.0, showing a purification factor of 1.35. However, the highest partition coefficient (14.42) was observed within a 26% (w/w) PEG 400 (g/mol) and a 20% (w/w) sodium citrate at pH 6.0. The independent variables which more influenced on the partition coefficient and recovery were citrate concentration and PEG mass molar, respectively. The results suggest that PEG–citrate ATPS is an interesting and efficient alternative to partition and recovery of phytase from fermentation broth produced by Absidia blakesleeana URM5604.
ISSN:0378-3812
1879-0224
DOI:10.1016/j.fluid.2012.01.004