Recovery and Purity of High Molar Mass Bio-hyaluronic Acid Via Precipitation Strategies Modulated by pH and Sodium Chloride

Author(s)

Cavalcanti, Andre D. D. ^[1] ; Melo, Bruna A. G. ^[1] ; Oliveira, Rhelvis C. ^[1] ; Santana, Maria H. A. ^[1]

Affiliation

^[1] Univ Estadual Campinas, Sch Chem Engn, Dev Biotechnol Microbial Proc Lab, Av Albert Einstein 00, BR-13083852 Campinas, SP - Brazil

• Document type: Journal article
• Source: Applied Biochemistry and Biotechnology; v. 188, n. 2, p. 527-539, JUN 2019.
• Web of Science Citations: 0

Abstract

The effects of ethanol/broth proportions and the number of steps at varying pH in the presence or absence of sodium chloride (NaCl) were studied as precipitation strategies for the recovery and purification of high molar mass bio-hyaluronic acid (Bio-HA). Bio-HA was synthesized by Streptococcus zooepidemicus in a culture medium containing glucose and soy peptones. A single-step precipitation was more attractive than multistep precipitation in terms of recovery and purity as well as decreased use of ethanol. The best conditions in the absence and presence of salt were 2:1 ethanol/broth (v/v) at pH4 (55.0 +/- 0.2% purity and 85.0 +/- 0.7% recovery) and 2:1 ethanol/broth (v/v) at pH7+2molL(-1) NaCl (59.0 +/- 0.9% purity and 82.0 +/- 4.3% recovery). Dynamic light scattering (DLS) spectra showed different particle sizes as a consequence of the changes in the molecular structure of HA, mainly with changes in pH. Although slight changes in distribution were observed, the average HA molar mass was not affected by the precipitation strategy, remaining on the order of 10(5)Da. Therefore, pH and NaCl modulated the precipitation performance of HA. These findings are relevant to further optimizing the precipitation step, thus minimizing costs in the later stages of HA purification. (AU)

• FAPESP's process: 15/23134-8 - Influence of the association between platelet-rich plasma and hyaluronic acid on the release of growth factors, cellular proliferation and osteogenic/ chondrogenic differentiation
• Grantee: Bruna Alice Gomes de Melo
• Support Opportunities: Scholarships in Brazil - Doctorate