Effect of Process Variables on Supercritical Fluid Disruption of Ralstonia eutropha Cells for Poly(R-hydroxybutyrate) Recovery

• Published in: Biotechnology progress Vol. 20; no. 6; pp. 1757 - 1765
• Main Authors: Khosravi-Darani, Kianoosh, Vasheghani-Farahani, Ebrahim, Shojaosadati, Seyed Abbas, Yamini, Yadollah
• Format: Journal Article
• Language: English
• Published: USA American Chemical Society 01.11.2004; American Institute of Chemical Engineers
• Subjects: Biological and medical sciences; Biotechnology; Cell Culture Techniques - methods; Cell Fractionation - methods; Cell Proliferation; Chromatography, Supercritical Fluid - methods; Cupriavidus necator - cytology; Cupriavidus necator - metabolism; Factor Analysis, Statistical; Fundamental and applied biological sciences. Psychology; Hydroxybutyrates - isolation & purification; Hydroxybutyrates - metabolism; Polyesters - isolation & purification; Polyesters - metabolism; Pressure; Protons; Ralstonia eutropha; Temperature; Ralstonia eutropha; Bacteria; Butyrate(hydroxy)polymer; Polymer; Supercritical state; Operating process; Recovery
• ISSN: 8756-7938; 1520-6033
• DOI: 10.1021/bp0498037

This research focuses on the disruption of the Gram‐negative bacterium Ralstonia eutropha cells by supercritical CO2 for poly( R‐hydroxybutyrate) (PHB) recovery. The variables affecting cell disruption such as drying strategy, type of modifier, and cultivation time, as well as operating pressure, temperature, and repeated release of supercritical CO2 pressure, have been studied. Effect of this disruption technique on PHB molecular mass was also investigated. PHB recovery was examined using a combination of this method and chemical pretreatments. For salt pretreatment, the cells were exposed to 140 mM NaCl and heat (60 °C, 1 h). The cells were also exposed to 0.2–0.8% (w/w) NaOH to examine the effect of alkaline pretreatment. Bacterial cells treated in growth phase exhibited less resistance to disruption than nutrient‐limited cells in the stationary phase. It was also found that the wet cells could be utilized to recover PHB, but purity of the product was lower than that obtained from freeze‐dried cells. Pretreatment with a minimum of 0.4% (w/w) NaOH was necessary to enable complete disruption with two times pressure release. Salt pretreatment was less effective; however, disruption was improved by the application of alkaline shock. The proposed method is economic and comparable with other recovery methods in terms of the percentage of PHB recovery and energy consumption, while it is environmentally more benign.