Extraction and purification of medium-chain-length polyhydroxyalkanoates produced by Streptomyces sp. using enzymes and an aqueous two-phase system
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Abstract
Polyhydroxyalkanoates (PHAs) are biodegradable polymers synthesized by bacteria, exhibiting properties similar to
petrochemical-based plastics while being environmentally friendly. Traditional extraction methods pose environmental
hazards, necessitating cost-effective and eco-friendly alternatives. This study aimed to investigate the extraction and purification of PHA from bacterial cells using enzymes and an aqueous two-phase system (ATPS). The bacteria used in this study were isolated from agricultural soil and cultivated in a nitrogen-limited minimal salts medium without complex carbon sources. Three enzymes—lysozyme, papain, and lipase—at concentrations ranging from 2 to 10 g/g cell, were employed to digest the bacterial cell wall. Subsequently, the purification of PHA was examined using five different ATPSs, and the composition of the extracted PHA was analyzed by gas chromatography-mass spectrometry (GC-MS). The results indicated that the isolated bacterium was identified as the Gram-positive species Streptomyces griseoincarnatus. Among the tested enzymes, papain at 10 g/g cell exhibited the highest efficiency in cell wall digestion, followed by lysozyme and lipase at the same concentration, with relative absorbance reductions of 81.9%, 34.2%, and 26.2%, respectively. The optimal extraction efficiency was achieved when lysozyme (10 g/g cell) was used to pre-digest the cell wall for 8 hours, followed by papain (10 g/g cell) for an additional 12 hours, yielding the highest PHA recovery of 87.2%. The PEG6000/K₂HPO₄ system was found to be the most effective ATPS, increasing the purity of the extracted PHA by 58.8%. The extracted PHA was identified as a medium-chain-length polymer. Therefore, the development of an enzymatic extraction method combined with ATPS purification provides an efficient and environmentally friendly approach for recovering PHA from bacterial cells.
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References
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