Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Aug;61(8):3113–3118. doi: 10.1128/aem.61.8.3113-3118.1995

Substrate specificities of bacterial polyhydroxyalkanoate depolymerases and lipases: bacterial lipases hydrolyze poly(omega-hydroxyalkanoates).

K E Jaeger 1, A Steinbüchel 1, D Jendrossek 1
PMCID: PMC167586  PMID: 7487042

Abstract

The substrate specificities of extracellular lipases purified from Bacillus subtilis, Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas fluorescens, and Burkholderia cepacia (former Pseudomonas cepacia) and of extracellular polyhydroxyalkanoate (PHA) depolymerases purified from Comamonas sp., Pseudomonas lemoignei, and P. fluorescens GK13, as well as that of an esterase purified from P. fluorescens GK 13, to various polyesters and to lipase substrates were analyzed. All lipases and the esterase of P. fluorescens GK13 but none of the PHA depolymerases tested hydrolyzed triolein, thereby confirming a functional difference between lipases and PHA depolymerases. However, most lipases were able to hydrolyze polyesters consisting of an omega-hydroxyalkanoic acid such as poly(6-hydroxyhedxanoate) or poly(4-hydroxybutyrate). The dimeric ester of hydroxyhexanoate was the main product of enzymatic hydrolysis of polycaprolactone by P. aeruginosa lipase. Polyesters containing side chains in the polymer backbone such as poly (3-hydroxybutyrate) and other poly(3-hydroxyalkanoates) were not or were only slightly hydrolyzed by the lipases tested.

Full Text

The Full Text of this article is available as a PDF (347.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson A. J., Dawes E. A. Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev. 1990 Dec;54(4):450–472. doi: 10.1128/mr.54.4.450-472.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandl H., Gross R. A., Lenz R. W., Fuller R. C. Pseudomonas oleovorans as a Source of Poly(beta-Hydroxyalkanoates) for Potential Applications as Biodegradable Polyesters. Appl Environ Microbiol. 1988 Aug;54(8):1977–1982. doi: 10.1128/aem.54.8.1977-1982.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Briese B. H., Jendrossek D., Schlegel H. G. Degradation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by aerobic sewage sludge. FEMS Microbiol Lett. 1994 Mar 15;117(1):107–111. doi: 10.1111/j.1574-6968.1994.tb06750.x. [DOI] [PubMed] [Google Scholar]
  4. CHOWDHURY A. A. POLY-BETA-HYDROXYBUTTERSAEURE ABBAUENDE BAKTERIEN UND EXOENZYM. Arch Mikrobiol. 1963 Dec 10;47:167–200. [PubMed] [Google Scholar]
  5. Delafield F. P., Doudoroff M., Palleroni N. J., Lusty C. J., Contopoulos R. Decomposition of poly-beta-hydroxybutyrate by pseudomonads. J Bacteriol. 1965 Nov;90(5):1455–1466. doi: 10.1128/jb.90.5.1455-1466.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Derewenda Z. S., Sharp A. M. News from the interface: the molecular structures of triacylglyceride lipases. Trends Biochem Sci. 1993 Jan;18(1):20–25. doi: 10.1016/0968-0004(93)90082-x. [DOI] [PubMed] [Google Scholar]
  7. FORSYTH W. G., HAYWARD A. C., ROBERTS J. B. Occurrence of poly-beta-hydroxybutyric acid in aerobic gram-negative bacteria. Nature. 1958 Sep 20;182(4638):800–801. doi: 10.1038/182800a0. [DOI] [PubMed] [Google Scholar]
  8. Haywood G. W., Anderson A. J., Ewing D. F., Dawes E. A. Accumulation of a Polyhydroxyalkanoate Containing Primarily 3-Hydroxydecanoate from Simple Carbohydrate Substrates by Pseudomonas sp. Strain NCIMB 40135. Appl Environ Microbiol. 1990 Nov;56(11):3354–3359. doi: 10.1128/aem.56.11.3354-3359.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jaeger K. E., Adrian F. J., Meyer H. E., Hancock R. E., Winkler U. K. Extracellular lipase from Pseudomonas aeruginosa is an amphiphilic protein. Biochim Biophys Acta. 1992 Apr 17;1120(3):315–321. doi: 10.1016/0167-4838(92)90254-b. [DOI] [PubMed] [Google Scholar]
  10. Jaeger K. E., Ransac S., Dijkstra B. W., Colson C., van Heuvel M., Misset O. Bacterial lipases. FEMS Microbiol Rev. 1994 Sep;15(1):29–63. doi: 10.1111/j.1574-6976.1994.tb00121.x. [DOI] [PubMed] [Google Scholar]
  11. Jendrossek D., Frisse A., Behrends A., Andermann M., Kratzin H. D., Stanislawski T., Schlegel H. G. Biochemical and molecular characterization of the Pseudomonas lemoignei polyhydroxyalkanoate depolymerase system. J Bacteriol. 1995 Feb;177(3):596–607. doi: 10.1128/jb.177.3.596-607.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jendrossek D., Müller B., Schlegel H. G. Cloning and characterization of the poly(hydroxyalkanoic acid)-depolymerase gene locus, phaZ1, of Pseudomonas lemoignei and its gene product. Eur J Biochem. 1993 Dec 1;218(2):701–710. doi: 10.1111/j.1432-1033.1993.tb18424.x. [DOI] [PubMed] [Google Scholar]
  13. Kouker G., Jaeger K. E. Specific and sensitive plate assay for bacterial lipases. Appl Environ Microbiol. 1987 Jan;53(1):211–213. doi: 10.1128/aem.53.1.211-213.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lee Y. P., Chung G. H., Rhee J. S. Purification and characterization of Pseudomonas fluorescens SIK W1 lipase expressed in Escherichia coli. Biochim Biophys Acta. 1993 Aug 11;1169(2):156–164. doi: 10.1016/0005-2760(93)90200-s. [DOI] [PubMed] [Google Scholar]
  16. Lesuisse E., Schanck K., Colson C. Purification and preliminary characterization of the extracellular lipase of Bacillus subtilis 168, an extremely basic pH-tolerant enzyme. Eur J Biochem. 1993 Aug 15;216(1):155–160. doi: 10.1111/j.1432-1033.1993.tb18127.x. [DOI] [PubMed] [Google Scholar]
  17. Mergaert J., Webb A., Anderson C., Wouters A., Swings J. Microbial degradation of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in soils. Appl Environ Microbiol. 1993 Oct;59(10):3233–3238. doi: 10.1128/aem.59.10.3233-3238.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Misset O., Gerritse G., Jaeger K. E., Winkler U., Colson C., Schanck K., Lesuisse E., Dartois V., Blaauw M., Ransac S. The structure-function relationship of the lipases from Pseudomonas aeruginosa and Bacillus subtilis. Protein Eng. 1994 Apr;7(4):523–529. doi: 10.1093/protein/7.4.523. [DOI] [PubMed] [Google Scholar]
  19. Ollis D. L., Cheah E., Cygler M., Dijkstra B., Frolow F., Franken S. M., Harel M., Remington S. J., Silman I., Schrag J. The alpha/beta hydrolase fold. Protein Eng. 1992 Apr;5(3):197–211. doi: 10.1093/protein/5.3.197. [DOI] [PubMed] [Google Scholar]
  20. SCHLEGEL H. G., GOTTSCHALK G., VON BARTHA R. Formation and utilization of poly-beta-hydroxybutyric acid by Knallgas bacteria (Hydrogenomonas). Nature. 1961 Jul 29;191:463–465. doi: 10.1038/191463a0. [DOI] [PubMed] [Google Scholar]
  21. SCHLEGEL H. G., KALTWASSER H., GOTTSCHALK G. [A submersion method for culture of hydrogen-oxidizing bacteria: growth physiological studies]. Arch Mikrobiol. 1961;38:209–222. [PubMed] [Google Scholar]
  22. Saito T., Suzuki K., Yamamoto J., Fukui T., Miwa K., Tomita K., Nakanishi S., Odani S., Suzuki J., Ishikawa K. Cloning, nucleotide sequence, and expression in Escherichia coli of the gene for poly(3-hydroxybutyrate) depolymerase from Alcaligenes faecalis. J Bacteriol. 1989 Jan;171(1):184–189. doi: 10.1128/jb.171.1.184-189.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schirmer A., Jendrossek D. Molecular characterization of the extracellular poly(3-hydroxyoctanoic acid) [P(3HO)] depolymerase gene of Pseudomonas fluorescens GK13 and of its gene product. J Bacteriol. 1994 Nov;176(22):7065–7073. doi: 10.1128/jb.176.22.7065-7073.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schirmer A., Jendrossek D., Schlegel H. G. Degradation of poly(3-hydroxyoctanoic acid) [P(3HO)] by bacteria: purification and properties of a P(3HO) depolymerase from Pseudomonas fluorescens GK13. Appl Environ Microbiol. 1993 Apr;59(4):1220–1227. doi: 10.1128/aem.59.4.1220-1227.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Steinbüchel A., Hustede E., Liebergesell M., Pieper U., Timm A., Valentin H. Molecular basis for biosynthesis and accumulation of polyhydroxyalkanoic acids in bacteria. FEMS Microbiol Rev. 1992 Dec;9(2-4):217–230. doi: 10.1111/j.1574-6968.1992.tb05841.x. [DOI] [PubMed] [Google Scholar]
  26. Stuer W., Jaeger K. E., Winkler U. K. Purification of extracellular lipase from Pseudomonas aeruginosa. J Bacteriol. 1986 Dec;168(3):1070–1074. doi: 10.1128/jb.168.3.1070-1074.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Timm A., Steinbüchel A. Formation of polyesters consisting of medium-chain-length 3-hydroxyalkanoic acids from gluconate by Pseudomonas aeruginosa and other fluorescent pseudomonads. Appl Environ Microbiol. 1990 Nov;56(11):3360–3367. doi: 10.1128/aem.56.11.3360-3367.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Winkler U. K., Stuckmann M. Glycogen, hyaluronate, and some other polysaccharides greatly enhance the formation of exolipase by Serratia marcescens. J Bacteriol. 1979 Jun;138(3):663–670. doi: 10.1128/jb.138.3.663-670.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. de Smet M. J., Eggink G., Witholt B., Kingma J., Wynberg H. Characterization of intracellular inclusions formed by Pseudomonas oleovorans during growth on octane. J Bacteriol. 1983 May;154(2):870–878. doi: 10.1128/jb.154.2.870-878.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES