Platform design for extraction and isolation of Bromelain: Complex formation and precipitation with carrageenan. (March 2017)

• Record Type: Journal Article
• Authors: Campos, Débora A.; Woitovich Valetti, Nadia; Oliveira, Ana; Pastrana-Castro, Lorenzo M.; Teixeira, José A.; Pintado, Maria Manuela; Picó, Guillermo
• Is Part Of: Process biochemistry. Volume 54(2017)
• Page Start: 156
• Page End: 161
• Publication Date: 2017-03
• Subjects: BR bromelain -- Carr carrageenan; Protein complex -- Enzyme mechanism -- Carrageenan -- Polyelectrolytes -- Bromelain; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2016.12.014
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Use of natural polysaccharides as precipitant agents of proteins. Formation of non-soluble columbic complexes. Recovery of proteins with maintenance of bioactivities;. Isolation, purification and recovery of BR, with 80% of yield of activity. Biological precipitation showed advantages when compared with traditional methods. Abstract: The main objective of this work was to investigate for the first time the molecular mechanism of complex formation between bromelain (a positively charged enzyme) and carrageenan (a natural strong polyelectrolyte, negatively charged) using spectroscopy techniques and thermodynamic approaches. The Bromelain-Carrageenan complex showed a maximal non-solubility at pH around 5.1. The solubility was dependent on pH and ionic strength of the medium. To re-dissolve the formed complex, the pH was changed and 500 mM of NaCl was added to the initial solution, proving the columbic mechanism for the formation of non-soluble complex. The formation of the carrageenan-bromelain complex increased in 8 °C the enzyme thermal stability, while its biological activity was not modified. The amount of total enzyme recovered in solution after precipitation with around 0.08% w/v of carrageenan was 85–90%.