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Abstract

Foam fractionation process for concentration of laccases from two Basidiomycete strains under different process conditions was investigated. Culture supernatants of Cerrena unicolor and Pleurotus sapidus containing active laccase were used with and without surfactant additives. Two surfactants: cationic cetrimonium bromide (CTAB) and non-ionic Polysorbate 80 were applied in the range from 0.2 mM to 1.5 mM. The pH levels ranging from 3 to 10 were examined with particular attention to pH=4, which is close to the pI of the enzymes. Results show that the source of the enzyme is significant in terms of partitioning efficiency in a foam fractionation process. Laccase from Cerrena unicolor showed the best activity partitioning coefficients between foamate and retentate of almost 200 with yields reaching 50% for pH 7.5 and concentration of CTAB cCTAB = 0.5 mM, whereas laccase from Pleurotus sapidus showed partitioning coefficients of up to 8 with 25% yield for pH 4 and cCTAB = 0.5 mM.

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Authors and Affiliations

Michał Blatkiewicz
Stanisław Ledakowicz
Anna Antecka
Andrzej Górak
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Abstract

The use of foam fractionation followed by aqueous two-phase extraction has emerged as a potential alternative to traditional liquid chromatography, hitherto irreplaceable in the purification of phycobiliproteins. The crude extracts of C-phycocyanin and allophycocyanin were obtained after Thermosynechococcus PCC 6715 biomass disintegration. The FF process with air flow of 2.4 L·h -1 resulted in purification factors up to 1.47 and partitioning coefficients of about 39, and did not require the addition of surfactants. A temperature of 35˚C allowed for the highest partitioning coefficient of 67.6 and yield of 76%; however, the purity of C-PC in condensate at this temperature was lower than at 25˚C. ATPE was tested in 20 different systems consisting of polyethylene glycol and phosphate or citrate salts, of which PEG1500-citrate gave the highest purification factor value of 2.31. Conversely, a partitioning coefficient of 2416 and 1094 were obtained for the PEG1500-phosphate and PEG3000-phosphate systems, respectively. Interestingly, the use of FF condensate in subsequent ATPE step resulted, for the first time, in the separation of the polymer phase into two fractions, one contained C-phycocyanin and the other allophycocyanin. It can be concluded that the use of a two-step system of FF and ATPE is a viable way to separate phycobiliproteins.
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Authors and Affiliations

Anna Antecka
1
ORCID: ORCID
Rafał Szeląg
1
Stanisław Ledakowicz
1
ORCID: ORCID

  1. Lodz University of Technology, Faculty of Process and Environmental Engineering, Department of Bioprocess Engineering, Wolczanska 213, 93-005 Lodz, Poland

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