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Thermotolerant poly(3-hydroxybutyrate)-degrading bacteria from hot compost and characterization of the PHB depolymerase of Schlegelella sp. KB1a

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Abstract

Eighteen gram-negative thermotolerant poly(3-hydroxybutyrate) (PHB)-degrading bacterial isolates (T max≈60°C) were obtained from compost. Isolates produced clearing zones on opaque PHB agar, indicating the presence of extracellular PHB depolymerases. Comparison of physiological characteristics and determination of 16S rRNA gene sequences of four selected isolates revealed a close relatedness of three isolates (SA8, SA1, and KA1) to each other and to Schlegelella thermodepolymerans and Caenibacterium thermophilum. The fourth strain, isolate KB1a, showed reduced similarities to the above-mentioned isolates and species and might represent a new species of Schlegelella. Evidence is provided that S. thermodepolymerans and C. thermophilum are only one species. The PHB depolymerase gene, phaZ, of isolate KB1a was cloned and functionally expressed in Escherichia coli. Purified PHB depolymerase was most active around pH 10 and 76°C. The DNA-deduced amino acid sequence of the mature protein (49.4 kDa) shared significant homologies to other extracellular PHB depolymerases with a domain substructure: catalytic domain type 2—linker domain fibronectin type 3—substrate-binding domain type 1. A catalytic triad consisting of S20, D104, and H138 and a pentapeptide sequence (GLS20AG) characteristic for PHB depolymerases (PHB depolymerase box, GLSXG) and for other serine hydrolases (lipase box, GXSXG) were identified.

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References

  • Altenbuchner J, Viell P, Pelletier I (1992) Positive selection vectors based on palindrome DNA sequences. Methods Enzymol 216:457–466

    Article  CAS  PubMed  Google Scholar 

  • Braaz R, Wong S-L, Jendrossek D (2002) Production of PHA depolymerase A (PhaZ5) from Paucimonas lemoignei in Bacillus subtilis. FEMS Microbiol Lett 209:237–241

    Article  CAS  PubMed  Google Scholar 

  • Chowdhury AA (1963) Poly-ß-hydroxybuttersäure abbauende Bakterien und Exoenzym. Arch Mikrobiol 47:167–200

    CAS  PubMed  Google Scholar 

  • Delafield FP, Doudoroff M, Palleroni NJ, Lusty CJ, Contopoulos R (1965) Decomposition of poly-β-hydroxybutyrate by pseudomonads. J Bacteriol 90:1455–1466

    CAS  PubMed  Google Scholar 

  • Doi Y, Steinbüchel A (2001) Biopolymers, vol 3A. Polyesters I—biological systems and biotechnological production. Wiley-VCH, Weinheim

  • Elbanna K, Lüdke-Eversloh T, van Trappen S, Mergaert J, Swings J, Steinbüchel A (2003) Schlegelella thermodepolymerans gen. nov., sp. nov., a novel thermophilic bacterium that degrades poly(3-hydroxybutyrate-co-3-mercaptopropionate). Int J Syst Evol Microbiol 53:1165–1168

    Article  CAS  PubMed  Google Scholar 

  • Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580

    CAS  PubMed  Google Scholar 

  • Handrick R, Reinhardt S, Focarete ML, Scandola M, Adamus G, Kowalczuk M, Jendrossek D (2001) A new type of thermoalkalophilic hydrolase of Paucimonas lemoignei with high specificity for amorphous polyesters of short-chain-length hydroxyalkanoic acids. J Biol Chem 276:36215–36224

    Article  CAS  PubMed  Google Scholar 

  • Handrick R, Reinhardt S, Jendrossek D (2004a) Purification, characterization and cloning of the “intracellular” PHB depolymerase of Rhodospirillum rubrum: an enzyme with high similarity to extracellular PHB depolymerases. Submitted for publication

  • Handrick R, Reinhardt S, Schultheiss D, Reichart T, Schüler D, Jendrossek V, Jendrossek D (2004b) Unraveling of the function of the Rhodospirillum rubrum activator of polyhydroxybutyrate (PHB) degradation: the activator is a PHB granule bound protein (phasin). J Bacteriol 186:2466–2475

    Article  CAS  PubMed  Google Scholar 

  • Haney PJ, Badger JH, Buldak GL, Reich CI, Woese CR, Olsen GJ (1999) Thermal adaptation analyzed by comparison of protein sequences from mesophilic and extremely thermophilic Methanococcus species. PNAS USA 30:3578–3583

    Google Scholar 

  • Jendrossek D (2002) Extracellular polyhydroxyalkanoate depolymerases: the key enzymes of PHA degradation. In: Doi Y, Steinbüchel A (eds) Biopolymers, vol 3B. Wiley-VCH, Weinheim, pp 47–77

  • Jendrossek D, Handrick R (2002) Microbial degradation of polyhydroxyalkanoates. Annu Rev Microbiol 56:403–432

    Google Scholar 

  • Jendrossek D, Knoke I, Habibian RB, Steinbüchel A, Schlegel HG (1993a) Degradation of poly(3-hydroxybutyrate), PHB, by bacteria and purification of a novel PHB depolymerase from Comamonas sp. J Environ Polym Degrad 1:53–63

    CAS  Google Scholar 

  • Jendrossek D, Müller B, Schlegel HG (1993b) Cloning and characterization of the poly(hydroxyalkanoic acid)-depolymerase gene locus, phaZ1, of Pseudomonas lemoignei and its gene product. Eur J Biochem 218:701–710

    CAS  PubMed  Google Scholar 

  • Jendrossek D, Backhaus M, Andermann M (1995a) Characterization of the extracellular poly(3-hydroxybutyrate) depolymerase of Comamonas sp. and of its structural gene. Can J Microbiol 41(Suppl 1):160–169

    CAS  PubMed  Google Scholar 

  • Jendrossek D, Frisse A, Behrends A, Andermann M, Kratzin HD, Stanislawski T, Schlegel HG (1995b) Biochemical and molecular characterization of the Pseudomonas lemoignei depolymerase system. J Bacteriol 177:596–607

    CAS  PubMed  Google Scholar 

  • Madison LL, Huisman GW (1999) Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic. Microbiol Mol Biol Rev 63:21–53

    CAS  PubMed  Google Scholar 

  • Manaia CM, Nunes OC, Nogales B (2003) Caenibacterium thermophilum gen. nov., sp. nov., isolated from a thermophilic aerobic digester of municipal sludge. Int J Syst Evol Microbiol 53:1375–1382

    Article  CAS  PubMed  Google Scholar 

  • Mergaert J, Swings J (1996) Biodiversity of microorganism that degrade bacterial and synthetic polyesters. J Ind Microbiol 17:463–469

    CAS  Google Scholar 

  • Merrick JM, Doudoroff M (1964) Depolymerization of poly-β-hydroxybutyrate by an intracellular enzyme system. J Bacteriol 88:60–71

    CAS  PubMed  Google Scholar 

  • Neumeier S (1994) Abbau thermoplastischer Biopolymere auf Poly-ß-Hydroxyalkanoat-Basis durch terrestrische and marine Pilze. Diploma thesis. Universität Regensburg

  • Nojiri M, Saito T (1997) Structure and function of poly(3-hydroxybutyrate) depolymerases from Alcaligenes faecalis T1. J Bacteriol 179:6965–6970

    CAS  PubMed  Google Scholar 

  • Schirmer A, Matz C, Jendrossek D (1995) Substrate specificities of PHA-degrading bacteria and active site studies on the extracellular poly(3-hydroxyoctanoic acid) [P(3HO)] depolymerase of Pseudomonas fluorescens GK13. Can J Microbiol 41(Suppl 1):170–179

    Google Scholar 

  • Shinohe T, Nojiri M, Saito T, Stanislawski T, Jendrossek D (1996) Determination of the active sites of the poly(3-hydroxybutyrate) depolymerase of Pseudomonas lemoignei (PhaZ5) and of Alcaligenes faecalis. FEMS Microbiol Lett 141:103–109

    Article  CAS  PubMed  Google Scholar 

  • Stinson MW, Merrick JM (1974) Extracellular enzyme secretion by Pseudomonas lemoignei. J Bacteriol 119:152–161

    CAS  PubMed  Google Scholar 

  • Takeda M, Koizumi J, Yabe K, Adachi K (1998) Thermostable poly(3-hydroxybutyrate) depolymerase of a thermophilic strain of Leptothrix sp. isolated from a hot spring. J Biosci Bioeng 85:375–380

    Article  CAS  Google Scholar 

  • Takeda M, Kitashima K, Adachi K, Hanaoka Y, Suzuki I, Koizumi J (2000) Cloning and expression of the gene encoding thermostable poly(3-hydroxybutyrate) depolymerase. J Biosci Bioeng 90:416–421

    CAS  Google Scholar 

  • Takeda M, Kamagata Y, Ghiorse WC, Hanada S, Koizumi J (2002) Caldimonas manganoxidans gen. nov., sp. nov., a poly(3-hydroxybutyrate)-degrading manganese-oxidizing thermophile. Int J Syst Evol Microbiol 52:895–900

    Article  CAS  PubMed  Google Scholar 

  • Tanio T, Fukui T, Shirakura Y, Saito T, Tomita K, Kaiho T (1982) An extracellular poly(3-hydroxybutyrate) depolymerase from Alcaligenes faecalis. Eur J Biochem 124:71–77

    CAS  PubMed  Google Scholar 

  • Yu Z, Mohn MM (1999) Isolation and characterization of thermophilic bacteria capable of degrading dehydroabietic acid. Can J Microbiol 45:513–519

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by the Deutsche Forschungsgemeinschaft. We thank Beate Müller (Umwelt Schutz Nord) for providing compost samples. Advice of the Deutsche Sammlung für Mikroorganismen und Zellkulturen (DSMZ) is greatfully acknowledged.

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  1. Institut für Mikrobiologie, Universität Stuttgart, Allmandring 31, 70550, Stuttgart, Germany

    Fabian Romen, Simone Reinhardt & Dieter Jendrossek

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  1. Fabian Romen
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  2. Simone Reinhardt
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  3. Dieter Jendrossek
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Correspondence to Dieter Jendrossek.

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This contribution is dedicated to Hans G. Schlegel in honor of his 80th birthday.

Fabian Romen and Simone Reinhardt share first authorship.

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Romen, F., Reinhardt, S. & Jendrossek, D. Thermotolerant poly(3-hydroxybutyrate)-degrading bacteria from hot compost and characterization of the PHB depolymerase of Schlegelella sp. KB1a. Arch Microbiol 182, 157–164 (2004). https://doi.org/10.1007/s00203-004-0684-2

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  • DOI: https://doi.org/10.1007/s00203-004-0684-2

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