Preparation and properties of bacteriophage-borne enzyme degrading bacterial exopolysaccharide

Bacteriophages infected different serotypes of Klebsiella were isolated from sewage. Among them, a heat-stable polysaccharide depolymerase enzyme which could degrade bacterial exopolysaccharide effectively was prepared from the phage infecting Klebsiella K13. Treatment at 60 ℃ for 30 min could inactivate most of the K13 phage, with the titration decreasing from 6.4×108 PFU/mL to 1.6×106 PFU/mL. However, no obvious loss of phage enzyme activity was found after this treatment. The optimum hydrolytic temperature of phage enzyme was 60 ℃, with an activity 57% higher than that at 30 ℃. The addition of phage enzyme could result in a rapid decrease of viscosity of exopolysaccharide (EPS) solution within minutes, indicating that K13 phage polysaccharide depolymerase acts as a kind of endo-glycanohydrolase. HPLC and reducing sugar analysis showed that the hydrolysis of EPS approached approximately the maximum at 4h when the final concentration of phage was 6.0 × 108 PFU/mL. The results showed that Klebsiella K13 phage depolymerase enzyme could be used as a good tool for the preparation of EPS oligosaccharide.