Abstract
Two polyhydroxyalkanoate depolymerases, PHAase I and PHAase II, were purified to homogeneity from the culture supernatant of an effective PHA-degrading bacterium, Pseudomonas mendocina DS04-T. The molecular masses of PHAase I and PHAase II were determined by SDS-PAGE as 59.4 and 33.8 kDa, respectively. Their optimum pH values were 8.5 and 8, respectively. Enzymatic activity was optimal at 50 °C. Both purified enzymes could degrade PHB, PHBV, and P(3HB-co-4HB). Addition of Na+ and K+ slightly increased the rate of PHAase II. EDTA significantly inhibited PHAase II but not PHAase I. Mercaptoethanol and H2O2 also inhibited the activities of both enzymes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Chanprateep S (2010) Current trends in biodegradable polyhydroxyalkanoates. J Biosci Bioeng 110:621–632
Jendrossek D, Handrick R (2002) Microbial degradation of polyhydroxyalkanoates. Annu Rev Microbiol 56:403–432
Jendrossek D, Schirmer A, Schlegel HG (1996) Biodegradation of polyhydroxyalkanoic acids. Appl Microbiol Biotechnol 46:451–463
Kasuya K, Inoue Y, Doi Y (1996) Adsorption kinetics of bacterial PHB depolymerase on the surface of polyhydroxyalkanoate films. Int J Biol Macromol 19:35–40
Kim DY, Rhee YH (2003) Biodergadation of microbial and synthetic polyesters by fungi. Appl Microbiol Biotechnol 61:300–308
Knoll M, Hamm TM, Wagner F, Martinez V, Pleiss J (2009) The PHA depolymerase engineering database: a systematic analysis tool for the diverse family of polyhydroxyalkanoate (PHA) depolymerases. BMC Bioinformatics 10:89–97
Madison LL, Huisman GW (1999) Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic. Microbiol Mol Biol Rev 63:21–53
Oda Y, Osaka H, Urakami T, Tonomura K (1997) Purification and properties of poly(3-hydroxybutyrate) depolymerase from the fungus Paecilomyces lilacinus D218. Curr Microbiol 34:230–232
Papaneophytou CP, Velali EE, Pantazaki AA (2011) Purification and characterization of an extracellular medium-chain length polyhydroxyalkanoate depolymerase from Thermus thermophilus HB8. Polym Degrad Stabil 96:670–678
Quinteros R, Goodwin S, Lenz RW, Park WH (1999) Extracellular degradation of medium chain length poly(beta-hydroxyalkanoates) by Comamonas sp. Int J Biol Macromol 25:135–143
Shirakura Y, Fukui T, Saito T, Okamoto Y, Narikawa T, Koide K, Tomita K, Takemasa T, Masamune S (1986) Degradation of poly(3-hydroxybutyrate) by poly(3-hydroxybutyrate) depolymerase from Alcaligenes faecalis T1. Biochim Biophys Acta 880:46–53
Takeda M, Koizumi JI, Yabe K, Adachi K (1998) Thermostable poly(3-hydroxybutyrate) depolymerase of a thermophilic strain of Leptothrix sp. isolated from a hot spring. J Ferment Bioeng 85:375–380
Tokiwa Y, Calabia BP (2004) Degradation of microbial polyesters. Biotechnol Lett 26:1181–1189
Wang ZY, Gao J, Li LL, Jiang HS (2012) Purification and characterization of an extracellular poly(3-hydroxybutyrate-co-3-hydroxyvalerate) depolymerase from Acidovorax sp. HB01. World J Microbiol Biotechnol 28:2395–2402
Acknowledgments
This work was supported by National Natural Science Foundation of China (Grant No. 31100099) and Science Project of Liaoning Province Education Office (L2011060).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Mao, H., Jiang, H., Su, T. et al. Purification and characterization of two extracellular polyhydroxyalkanoate depolymerases from Pseudomonas mendocina . Biotechnol Lett 35, 1919–1924 (2013). https://doi.org/10.1007/s10529-013-1288-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-013-1288-1