Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/244242
Type: Artigo de periódico
Title: Liquid-liquid Extraction Of Lipase Produced By Psychrotrophic Yeast Leucosporidium Scottii L117 Using Aqueous Two-phase Systems
Author: Fernandes Duarte
Alysson Wagner; Lopes
Andre Moreni; Dutra Molino
Joao Vitor; Pessoa
Adalberto; Sette
Lara Duraes
Abstract: Aqueous two-phase systems (ATPS) have been used in biomolecules separation and as an efficient alternative to traditional purification systems for lipases extraction. Here, we investigated the partitioning and recovery of lipase derived from Leucosporidium scottii L117 using ATPS and aqueous two-phase micellar systems (ATPMS). Thus, we evaluated three ATPS: (i) polyethylene glycol (PEG)/phosphate salts and (ii) PEG/polyacrylic acid (NaPA) in different molecular weights (1500, 4000 and 8000 g/mol). (iii) Triton X-114 (TX-114)/McIlvaine buffer pH 7.0 in different conditions (2.0% (w/w) of TX-114 at 25.0 and 28.0 degrees C). The PEG/phosphate and PEG/NaPA systems resulted in a great loss of enzymatic activity; thus these systems do not represent viable alternatives for these lipase extraction. The micellar systems yielded the best results for lipase extraction with enzyme activity balances ranging between 84.7% and 113.05%. After optimizing the micellar system by experimental design of the partition coefficient of lipase increased by 10.3-fold (0.75-7.76). Lipase preferentially partitioned into the micelle-rich phase with K-Lip = 7.76, % RECBot = 93.85% and PF = 1.2 at 25.03 degrees C, 5.1 pH and 10.38% TX-114 and K-Lip, = 4.77, % RECBot = 73.53% and PF = 1.97 at 28.00 degrees C, 4.5 pH and 8.0% TX-114, indicating that the ATPMS represents an alternative to purification/extraction of lipase L scottii L117. A crude lipase extract was also evaluated to define the optimum pH and temperature. Lipase reached optimal activity at 40 degrees C, and remained stable in pH values ranging from pH 3.0 to 8.0 and temperatures from 20.0 to 45.0 degrees C, with relative residual lipase activity above 80% after 30 min of incubation. (C) 2015 Elsevier B.V. All rights reserved.
Subject: Biochemical-characterization
Microbial Lipases
Micellar Systems
Phospholipase-b
Ionic Liquids
Purification
Protein
Cloning
Biotechnology
Performance
Country: AMSTERDAM
Editor: ELSEVIER SCIENCE BV
Rights: embargo
Identifier DOI: 10.1016/j.seppur.2015.10.001
Address: http://www.sciencedirect.com/science/article/pii/S1383586615302525
Date Issue: 2015
Appears in Collections:Unicamp - Artigos e Outros Documentos

Files in This Item:
File SizeFormat 
wos_000366537900014.pdf3.5 MBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.