Enzymatic characterization of a depolymerase from the isolated bacterium Variovorax sp. C34 that degrades poly(enriched lactate-co-3-hydroxybutyrate)

Poly[(R)-lactate-co-(R)-3-hydroxybutyrate] [P(LA-co-3HB)] is a biobased polyester with semitransparent and flexible properties produced in engineered bacteria carrying an LA-polymerizing enzyme. In this study, we attempted to isolate the P(enriched LA-co-3HB)-degrading bacteria from soil samples in order to identify enzymes with the capacity to degrade this new type of polymer. Among approximately 500 samples, the Gram-negative bacterium C34, which exhibited potent P(enriched LA-co-3HB)-degrading activity, was isolated based on the decrease in the turbidity of culture medium supplemented with emulsified P(67 mol% LA-co-3HB). Based on its 16S rDNA sequence, this isolated bacterium was identified as a member of Variovorax sp.. Next, we attempted to isolate and purify the depolymerase that contributes to the polymer degradation from the culture supernatant of strain C34. The purified enzyme had a molecular mass of 42 kDa and exhibited degradation activity towards the P(67 mol% LA-co-3HB) as well as 3HB homopolymer [P(3HB)], but not LA homopolymers (PDLA and PLLA). On the other hand, a well-characterized depolymerase of P(3HB) derived from Alcaligenes faecalis T1 did not degrade P(67 mol% LA-co-3HB), PDLA or PLLA. This result suggests that the newly isolated depolymerase differs from the P(3HB) depolymerase from A. faecalis.