Abstract
Strain DSDY0501 with polyhydroxybutyrate (PHB)-degrading ability was isolated from activated sludge. The morphological, physiological, and biochemical properties of the strain and phylogenetic analysis indicated that the strain belongs to Pseudomonas sp. The strain used PHB as the sole carbon source and degraded PHB films completely in 21 h in liquid culture. An extracellular PHB depolymerase was purified from the supernatant of the culture by ultrafiltration and Sephacryl S-200 gel filtration. The specific activity of the purified enzyme increased 24.2-fold, and the recovery yield was 16.61%. Extracellular PHB depolymerase, a monomeric enzyme with a molecular weight of approximately 57.9 kDa, showed optimum activity at 60 °C and pH 9.0, and was stable in the temperature range of 10-60 °C and a pH range of 6.0-10.0. The secondary structure of the enzyme contained approximately 60% α-helix and 40% β-pleated sheet according to the circular dichroism spectrum. Mass spectrum analysis showed that the main degradation product of the enzyme was PHB monomer, indicating the exo-type action of this PHB depolymerase.