The Effectiveness of Polyhydroxyalkanoate (PHA) Extraction Methods in Gram-Negative Pseudomonas putida U.

Abstract

Bioplastics are emerging as a promising solution to reduce pollution caused by petroleum-based plastics. Among them, polyhydroxyalkanoates (PHAs) stand out as viable biotechnological alternatives, though their commercialization is limited by expensive downstream processes. Traditional PHA extraction methods often involve toxic solvents and high energy consumption, underscoring the need for more sustainable approaches. This study evaluated physical and chemical methods to extract PHAs from Pseudomonas putida U, a bacterium known to produce poly-3-hydroxyoctanoate P(3HO). Lyophilized cells underwent six extraction methods, including the use of the following: boiling, sonication, sodium hypochlorite (NaClO), sodium dodecyl sulfate (SDS), sodium hydroxide (NaOH), and chloroform. Physical methods such as boiling and sonication achieved yields of 70% and 60%, respectively, but P(3HO) recovery remained low (30-40%). NaClO extraction provided higher yields (80%) but resulted in significant impurities (70%). NaOH methods offered moderate yields (50-80%), with P(3HO) purities between 50% and 70%, depending on the conditions. Spectroscopic and analytical techniques (FTIR, TGA, NMR, GPC) identified 0.05 M NaOH at 60 °C as the optimal extraction condition, delivering high P(3HO) purity while minimizing environmental impact. This positions NaOH as a sustainable alternative to traditional halogenated solvents, paving the way for more eco-friendly PHA production processes.