Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Peculiarities of PHA granules preparation and PHA depolymerase activity determination

  • 788 Accesses

  • 39 Citations

Abstract

An extensive amount of knowledge on biochemistry of poly(3-hydroxyalkanoic acid) (PHA) synthesis and on its biodegradation has accumulated during the last two decades. Numerous genes encoding enzymes involved in the formation of PHA and in PHA degradation (PHA depolymerases) were cloned and characterized from many microorganisms. A large variety of methods exists for determination of PHA depolymerase activity and for preparation of the polymeric substrate (PHA). Unfortunately, results obtained with these different methods cannot be compared directly because they highly depend on the assay method applied and on the history of PHA granules preparation. In this contribution, the peculiarities, advantages, disadvantages and limitations of existing PHA depolymerase assay methods are described.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2

References

  1. Abe H, Madsubara I, Doi Y, Hori Y, Yamaguchi A (1994) Physical properties and enzymatic degradability of poly(3-hydroxybutyrate) stereoisomers with different stereoregularities. Macromolecules 27:6018–6025

  2. Abe T, Kobayashi T, Saito T (2005) Properties of a novel intracellular poly(3-hydroxybutyrate) depolymerase with high specific activity (PhaZd) in Wautersia eutropha H16. J Bacteriol 187:6982–6990

  3. Alper R, Lundgren DG, Marchessault RH, Cote WH (1963) Properties of poly-β-hydroxybutyrate. 1. General considerations concerning the naturally occurring polymer. Biopolymers 1:545–556

  4. Anderson AJ, Dawes EA (1990) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev 54:450–472

  5. Bachmann B, Seebach D (1999) Investigation of the enzymatic cleavage of diastereomeric oligo(3-hydroxybutanoates) containing two to eight HB units. A model for the stereoselectivity of PHB depolymerase from Alcaligenes faecalis T1. Macromolecules 32:1777–1784

  6. Behrends A, Klingbeil B, Jendrossek D (1996) Poly(3-hydroxybutyrate) depolymerases bind to their substrate by a C-terminal located substrate binding site. FEMS Microbiol Lett 143:191–194

  7. Boatman ES (1964) Observations on the fine structure of spheroplasts of Rhodospirillum rubrum. J Cell Biol 20:297–311

  8. Braaz R, Wong SL, Jendrossek D (2002) Production of PHA depolymerase A (PhaZ5) from Paucimonas lemoignei in Bacillus subtilis. FEMS Microbiol Lett 209:237

  9. Braaz R, Handrick R, Jendrossek D (2003) Identification and characterisation of the catalytic triad of the alkaliphilic thermotolerant PHA depolymerase PhaZ7 of Paucimonas lemoignei. FEMS Microbiol Lett 224:107–112

  10. Brandl H, Gross RA, Lenz RW, Fuller RC (1988) Pseudomonas oleovorans as a source of poly(beta-hydroxyalkanoates) for potential applications as biodegradable polyesters. Appl Environ Microbiol 54:1977–1982

  11. Briese BH, Jendrossek D, Schlegel HG (1994) Degradation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by aerobic sewage sludge. FEMS Microbiol Lett 117:107–111

  12. Calabia BP, Tokiwa Y (2006) A novel PHB depolymerase from a thermophilic Streptomyces sp. Biotechnol Lett 28:383–388

  13. Choi MH et al (2003) Biosynthesis and local sequence specific degradation of poly(3-hydroxyvalerate-co-4-hydroxybutyrate) in Hydrogenophaga pseudoflava. Biomacromolecules 4:38–45

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

  15. Cornibert J, Marchessault RH (1972) Physical properties of poly-β-hydroxybutyrate. IV. Conformational analysis and crystalline structure. J Mol Biol 71:735–756

  16. De Eugenio LI, Garcia P, Luengo JM, Sanz JM, San Roman J, Garcia JL, Prieto MA (2006) Biochemical evidence that phaZ gene encodes a specific intracellular medium chain length polyhydroxyalkanoate depolymerase in Pseudomonas putida KT2442: characterization of a paradigmatic enzyme. J Biol Chem, Dec 14 (in press)

  17. de Koning GJM, Lemstra PJ (1992) The amorphous state of poly[(R)-3-hydroxyalkanoate] in vivo. Polymer 33:3292–3294

  18. de Koning GJM et al (1994) A biodegradable rubber by cross-linking poly(hydroxyalkanoate) from Pseudomonas fluorescens. Polymer 35:2090–2097

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

  20. De Smet MJ, Eggink G, Witholt B (1983) Characterization of intracellular inclusions formed by Pseudomonas oleovorans during growth on octane. J Bacteriol 154:870–878

  21. Do YK, Hyun CK, Sun YK, Young HR (2005) Molecular characterization of extracellular medium-chain-length poly(3-hydroxyalkanoate) depolymerase genes from Pseudomonas alcaligenes strains. J Microbiol 43:285–294

  22. Doi Y, Katsuyuki M, Kasuya K, Yamada K (1994) Biodegradation of biosynthetic and chemosynthetic polyhydroxyalkanoates. In: Doi Y, Fukuda K (eds) Biodegradable plastics and polymers. Elsevier Science B.V., pp 39–51

  23. Elbanna K, Lütke-Eversloh T, Jendrossek D, Luftmann H, Steinbüchel A (2004) Studies on the biodegradability of polythioester copolymers and homopolymers by polyhydroxyalkanoate (PHA)-degrading bacteria and PHA depolymerases. Arch Microbiol 182:212–224

  24. Ellar D, Lundgren DG, Okamura K, Marchessault RH (1968) Morphology of poly-beta-hydroxybutyrate granules. J Mol Biol 35:489–502

  25. Foster LJR, Lenz RW, Fuller RC (1994) Quantitative determination of intracellular depolymerase activity in Pseudomonas oleovorans inclusions containing poly-3-hydroxyalkanoates with long alkyl substituents. FEMS Microbiol Lett 118:279

  26. Foster LJR, Stuart ES, Tehrani A, Lenz RW, Fuller RC (1996) Intracellular depolymerase and polyhydroxyoctanoate granule integrity in Pseudomonas oleovorans. Int J Biol Macromol 19:177–183

  27. Foster LJR, Lenz RW, Fuller RC (1999) Intracellular depolymerase activity in isolated inclusion bodies containing polyhydroxyalkanoates with long alkyl and functional substituents in the side chain. Int J Biol Macromol 26:187

  28. Fukui T, Yoshimoto A, Matsumoto M, Hosokawa S, Saito T (1976) Enzymatic synthesis of poly-beta-hydroxybutyrate in Zoogloea ramigera. Arch Microbiol 110:149–156

  29. Fuller RC, O’Donnell JP, Saulnier J, Redlinger TE, Foster J, Lenz RW (1992) The supramolecular architecture of the polyhydroxyalkanoate inclusions in Pseudomonas oleovorans. FEMS Microbiol Rev 103:279–288

  30. Gao D, Maehara A, Yamane T, Ueda S (2001) Identification of the intracellular polyhydroxyalkanoate depolymerase gene of Paracoccus denitrificans and some properties of the gene product. FEMS Microbiol Lett 196:159–164

  31. Gebauer B, Jendrossek D (2006) Assay of poly(3-hydroxybutyrate) depolymerase activity and product determination. Appl Environ Microbiol 72:6094–6100

  32. Gonda KE, Jendrossek D, Molitoris HP (2000) Fungal degradation of the thermoplastic polymer poly-β-hydroxybutyric acid (PHB) under simulated deep sea pressure. Hydrobiologia 426:173–183

  33. Griebel RJ, Merrick JM (1971) Metabolism of poly-beta-hydroxybutyrate: effect of mild alkaline extraction on native poly-β-hydroxybutyrate granules. J Bacteriol 108:782–789

  34. Griebel R, Smith Z, Merrick JM (1968) Metabolism of poly-beta-hydroxybutyrate. I. Purification, composition, and properties of native poly-beta-hydroxybutyrate granules from Bacillus megaterium. Biochemistry 7:3676–3681

  35. Handrick R, Reinhardt S, Jendrossek D (2000) Mobilization of poly(3-hydroxybutyrate) in Ralstonia eutropha. J Bacteriol 182:5916–5918

  36. Handrick R et al (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

  37. Handrick R, Reinhardt S, Kimmig P, Jendrossek D (2004a) The “intracellular” poly(3-hydroxybutyrate) (PHB) depolymerase of Rhodospirillum rubrum is a periplasm-located protein with specificity for native PHB and with structural similarity to extracellular PHB depolymerases. J Bacteriol 186:7243–7253

  38. Handrick R et al (2004b) Unraveling 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

  39. Handrick R, Technow U, Reichart T, Reinhardt S, Sander T, Jendrossek D (2004c) The activator of the Rhodospirillum rubrum PHB depolymerase is a polypeptide that is extremely resistant to high temperature (121 degrees C) and other physical or chemical stresses. FEMS Microbiol Lett 230:265–274

  40. Hippe H, Schlegel HG (1967) Hydrolyse von PHBS durch intracelluläre Depolymerase von Hydrogenomonas H16. Arch Mikrobiol 56:278–299

  41. Hisano T et al (2006) The crystal structure of polyhydroxybutyrate depolymerase from Penicillium funiculosum provides insights into the recognition and degradation of biopolyesters. J Mol Biol 356:993

  42. Horowitz DM, Sanders KM (1994) Amorphous, biomimetic granules of polyhydroxybutyrate: preparation, characterization, and biological implications. J Am Chem Soc 116:2695–2702

  43. Horowitz DM, Sanders JKM (1995) Biomimetic, amorphous granules of polyhydroxyalkanoates: composition, mobility, and stabilization in vitro by proteins. Can J Microbiol 41:115–123

  44. Horowitz DM, Brennan EM, Koon JJ, Gerngross TU (1999) Novel thermal route to an amorphous, film-forming polymer latex. Macromolecules 32:3347–3352

  45. Huisman GW, Wonink E, Meima R, Kazemier B, Terpstra P, Witholt B (1991) Metabolism of poly(3-hydroxyalkanoates) (PHAs) by Pseudomonas oleovorans. Identification and sequences of genes and function of the encoded proteins in the synthesis and degradation of PHA. J Biol Chem 266:2191–2198

  46. Jendrossek D (1998) Microbial degradation of polyesters: a review on extracellular poly(hydroxy alkanoic acid) depolymerases. Polym Degrad Stab 59:317–325

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

  48. 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

  49. 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

  50. Jendrossek D et al (1995) Biochemical and molecular characterization of the Pseudomonas lemoignei polyhydroxyalkanoate depolymerase system. J Bacteriol 177:596–607

  51. Jendrossek D, Schirmer A, Schlegel HG (1996) Biodegradation of polyhydroxyalkanoic acids. Appl Microbiol Biotechnol 46:451–463

  52. Jendrossek D, Schirmer A, Handrick R (1997) Recent advances in characterization of bacterial PHA depolymerase. In: Eggink G, Steinbüchel A, Poirier Y, Witholt B (eds) 1996 International symposium on bacterial polyhydroxyalkanoates. NRC Research, Ottawa, pp 89–101

  53. Jiang Y, Ye J, Wu H, Zhang H (2004) Cloning and expression of the polyhydroxyalkanote depolymerase gene from Pseudomonas putida, and characterization of the gene product. Biotechnol Lett 26:1585–1588

  54. Kadouri D, Jurkevitch E, Okon Y (2003) Poly β-hydroxybutyrate depolymerase (PhaZ) in Azospirillum brasilense and characterization of a phaZ mutant. Arch Microbiol 180:309–318

  55. Kawaguchi Y, Doi Y (1990) Structure of native poly(3-hydroxybutyrate) granules characterized by X-ray diffraction. FEMS Microbiol Lett 70:151–156

  56. Kikkawa Y, Yamashita K, Hiraishi T, Kanesato M, Doi Y (2005) Dynamic adsoption behavior of poly(3-hydroxybutyrate) depolymerase onto polyester surface investigated by QCM and AFM. Biomacromolecules 6:2084–2090

  57. Kim DY, Rhee YH (2003) Biodegradation of microbial and synthetic polyesters by fungi. Appl Microbiol Biotechnol 61:300–308

  58. Kim DY, Nam JS, Rhee YH (2002) Characterization of an extracellular medium-chain-length poly(3-hydroxyalkanoate) depolymerase from Pseudomonas alcaligenes LB19. Biomacromolecules 3:291–296

  59. Kobayashi T, Saito T (2003) Catalytic triad of intracellular poly(3-hydroxybutyrate) depolymerase (PhaZ1) in Ralstonia eutropha H16. J Biosci Bioeng 96:487–492

  60. Kobayashi T, Nishikori K, Saito T (2004) Properties of an intracellular poly(3-hydroxybutyrate) depolymerase (PhaZ1) from Rhodobacter spheroides. Curr Microbiol 49:199–202

  61. Kobayashi T, Uchino K, Abe T, Yamazaki Y, Saito T (2005) Novel intracellular 3-hydroxybutyrate-oligomer hydrolase in Wautersia eutropha H16. J Bacteriol 187:5129–5135

  62. Lee SY, Lee Y (2003) Metabolic engineering of Escherichia coli for production of enantiomerically pure (R)-(−)-hydroxycarboxylic acids. Appl Environ Microbiol 69:3421–3426

  63. Lundgren DG, Pfister RM, Merrick JM (1964) Structure of poly-beta-hydroxybutyric acid granules. J Gen Microbiol 34:441–446

  64. Lundgren DG, Alper R, Schnaitman C, Marchessault RH (1965) Characterization of poly-beta-hydroxybutyrate extracted from different bacteria. J Bacteriol 89:245–251

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

  66. Marchessault RH, Saracovan I (1995) Artificial granules suspension of long side chain poly(3-hydroxyalkanoate). Can J Microbiol 41(Suppl 1):138–142

  67. Mayer F (1992) Structural aspects of poly-β-hydroxybutyrate granules. FEMS Microbiol Rev 103:265–268

  68. Mayer F, Madkour MH, Pieper-Fürst U, Wieczorek R, Liebergesell M, Steinbüchel A (1996) Electron microscopic observations on the macromolecular organization of the boundary layer of bacterial PHA inclusion bodies. J Gen Microbiol 42:445–455

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

  70. Merrick JM, Lundgren DG, Pfister RM (1965) Morphological changes in poly-beta-hydroxybutyrate granules associated with decreased susceptibility to enzymatic hydrolysis. J Bacteriol 89:234–239

  71. Merrick JM, Steger R, Dombroski D (1999) Hydrolysis of native poly(hydroxybutyrate) granules (PHB), crystalline PHB, and artificial amorphous PHB granules by intracellular and extracellular depolymerases. Int J Biol Macromol 25:129–134

  72. Molitoris HP, Moss ST, de Koning GJM, Jendrossek D (1996) Scanning electron microscopy of polyhydroxyalkanoate degradation by bacteria. Appl Microbiol Biotechnol 46:570–579

  73. Müller B, Jendrossek D (1993) Purification and properties of a poly(3-hydroxyvalerate) depolymerase from Pseudomonas lemoignei. Appl Microbiol Biotechnol 38:487–492

  74. Müller H-M, Seebach D (1993) Poly(hydroxyalkanoates): a fifth class of physiologically important organic biopolymers? Angewandte Chemie 32:477–502

  75. Nobes GAR, Marchessault RH, Chanzy H, Briese BH, Jendrossek D (1996) Splintering of poly(3-hydroxybutyrate) single crystals by PHB-depolymerase A from Pseudomonas lemoignei. Macromolecules 29:8330–8333

  76. Numata K, Kikkawa Y, Tsuge T, Iwata T, Doi Y, Abe H (2005) Enzymatic degradation processes of poly[(R)-3-hydroxybutyric acid] and poly[(R)-3-hydroxybutyric acid-co-(R)-3-hydroxyvaleric acid] single crystals revealed by atomic force microscopy: effects of molecular weight and second-monomer composition on erosion rates. Biomacromolecules 6:2008–2016

  77. Oh JS, Choi MH, Yoon SC (2005) In vivo 13C-NMR spectroscopic study of polyhydroxyalkanoic acid degradation kinetics in bacteria. J Microbiol Biotechnol 15:1330–1336

  78. Ohura T, Aoyagi Y, Takagi K, Yoshida Y, Kasuya K, Doi Y (1999) Biodegradation of poly(3-hydroxyalkanoic acids) fibers and isolation of poly(3-hydroxybutyric acid)-degrading microorganisms under aquatic environments. Polym Degrad Stab 63:23–29

  79. Pohlmann A et al (2006) Genome sequence of the bioplastic-producing “Knallgas” bacterium Ralstonia eutropha H16. Nat Biotechnol 24:1257–1262

  80. Pötter M, Steinbüchel A (2005) Poly(3-hydroxybutyrate) granule-associated proteins: impacts on poly(3-hydroxybutyrate) synthesis and degradation. Biomacromolecules 6:552–560

  81. Pötter M et al (2004) The complex structure of polyhydroxybutyrate (PHB) granules: four orthologous and paralogous phasins occur in Ralstonia eutropha. Microbiology 150:2301–2311

  82. Prieto MA, Bühler B, Jung K, Witholt B, Kessler B (1999) PhaF, a polyhydroxyalkanoate-granule-associated protein of Pseudomonas oleovorans GPo1 involved in the regulatory expression system for pha genes. J Bacteriol 181:858

  83. Ramsay BA, Saracovan I, Ramsay JA, Marchessault RH (1994) Method for the isolation of microorganisms producing extracellular long-side-chain poly(-hydroxyalkanoate) depolymerase. J Environ Polym Degrad 2:1–7

  84. Ren Q et al (2005) Bacterial poly(hydroxyalkanoates) as a source of chiral hydroxyalkanoic acids. Biomacromolecules 6:2290–2298

  85. Rhee YH, Kim YH, Shin KS (2006) Characterization of an extracellular poly(3-hydroxyoctanoate) depolymerase from the marine isolate, Pseudomonas luteola M13-4. Enzyme Microb Technol 38:529–535

  86. Romen F, Reinhardt S, Jendrossek D (2004) Thermotolerant poly(3-hydroxybutyrate)-degrading bacteria from hot compost and characterization of the PHB depolymerase of Schlegellella sp. KB1a. Arch Microbiol 182:157–164

  87. Saegusa H, Shiraki M, Kanai C, Saito T (2001) Cloning of an intracellular poly[d(−)-3-hydroxybutyrate] depolymerase gene from Ralstonia eutropha H16 and characterization of the gene product. J Bacteriol 183:94–100

  88. Saito T, Kobayashi T (2002) Intracellular degradation of PHAs. In: Doi Y, Steinbüchel A (eds) Biopolymers. Wiley-VCH, pp 23–40

  89. Saito T, Saegusa H, Miyata Y, Fukui T (1992) Intracellular degradation of poly(3-hydroxybutyrate) granules of Zoogloea ramigera I-16-M. FEMS Microbiol Rev 9:333–338

  90. Saito T, Takizawa K, Saegusa H (1995) Intracellular poly(3-hydroxybutyrate) depolymerase in Alcaligenes eutrophus. Can J Microbiol 41(Suppl 1):187–191

  91. Sandoval A et al (2005) Production of 3-hydroxy-n-phenylalkanoic acids by a genetically engineered strain of Pseudomonas putida. Appl Microbiol Biotechnol 67:97–105

  92. Scandola M, Pizzoli M, Ceccorulli G, Cesaro A, Paolletti S, Navarini L (1988) Viscoelastic and thermal properties of bacterial poly(-(−)-[beta]-hydroxybutyrate). Int J Biol Macromol 10:373–377

  93. Schirmer A, Jendrossek D (1994) Molecular characterization of the extracellular poly(3-hydroxyoctanoic acid) [P(3HO)] depolymerase gene of Pseudomonas fluorescens GK13 and of its gene product. J Bacteriol 176:7065–7073

  94. Schirmer A, Jendrossek D, Schlegel HG (1993) 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 59:1220–1227

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

  96. Schöber U, Thiel C, Jendrossek D (2000) Poly(3-hydroxyvalerate) depolymerase of Pseudomonas lemoignei. Appl Environ Microbiol 66:1385–1392

  97. Shuster CW, Doudoroff M (1962) A cold-sensitive d(−) beta-hydroxybutyric acid dehydrogenase from Rhodospirillum rubrum. J Biol Chem 237:603–607

  98. Spyros A, Kimmich R, Briese BH, Jendrossek D (1997) 1H NMR imaging study of enzymatic degradation of poly(3-hydroxybutyrate) and poly(3-hydroxyvalerate). Macromolecules 30:8218–8225

  99. Steinbüchel A, Hofrichter M (2001) Biopolymers. Wiley-VCH, Weinheim; Chichester

  100. Steinbüchel A et al (1995) Considerations on the structure and biochemistry of bacterial polyhydroxyalkanoic acid inclusions. Can J Microbiol 41(Suppl 1):94–105

  101. Stuart ES, Foster LJR, Lenz RW, Fuller RC (1996) Intracellular depolymerase functionality and location in Pseudomonas oleovorans inclusions containing polyhydroxyoctanoate. Int J Biol Macromol 19:171–176

  102. Stuart ES, Tehrani A, Valentin HE, Dennis D, Lenz RW, Fuller RC (1998) Protein organization on the PHA inclusion cytoplasmic boundary. J Biotechnol 64:137

  103. Taidi B, Mansfield D, Anderson AJ (1995) Turnover of poly(3-hydroxybutyrate) (PHB) and its influence on the molecular mass of the polymer accumulated by Alcaligenes eutrophus during batch culture. FEMS Microbiol Lett 129:201–206

  104. Tanaka Y, Saito T, Fukui T, Tanio T, Tomita K (1981) Purification and properties of d(−)-3-hydroxybutyrate-dimer hydrolase from Zoogloea ramigera I-16-M. Eur J Biochem 118:177–182

  105. Tanio T et al (1982) An extracellular poly(3-hydroxybutyrate) depolymerase from Alcaligenes faecalis. Eur J Biochem 124:71–77

  106. Timm A, Steinbüchel A (1990) Formation of polyesters consisting of medium-chain-length 3-hydroxyalkanoic acids from gluconate by Pseudomonas aeruginosa and other fluorescent pseudomonads. Appl Environ Microbiol 56:3360–3367

  107. Tomasi G, Scandola M, Briese BH, Jendrossek D (1996) Enzymatic degradation of bacterial poly(3-hydroxybutyrate) by a depolymerase from Pseudomonas lemoignei. Macromolecules 29:507–513

  108. Tseng CL, Chen HJ, Shaw GC (2006) Identification and characterization of the Bacillus thuringiensis phaZ gene, encoding new intracellular poly-3-hydroxybutyrate depolymerase. J Bacteriol 188:7592–7599

  109. Uefuji M, Ken-ichi K, Doi Y (1997) Enzymatic degradation of poly[(R)-3-hydroxybutyrate]: secretion and properties of PHB depolymerase from Pseudomonas stutzeri. Polym Degrad Stab 58:275–281

  110. Wieczorek R, Pries A, Steinbüchel A, Mayer F (1995) Analysis of a 24-kilodalton protein associated with the polyhydroxyalkanoic acid granules in Alcaligenes eutrophus. J Bacteriol 177:2425–2435

  111. Wieczorek R, Steinbüchel A, Schmidt B (1996) Occurrence of polyhydroxyalkanoic acid granule-associated proteins related to the Alcaligenes eutrophus H16 GA24 protein in other bacteria. FEMS Microbiol Lett 135:23–30

  112. Williamson DH, Wilkinson JF (1958) The isolation and estimation of the β-hydroxybutryrate inclusions of Bacillus species. J Gen Microbiol 19:198–209

  113. Yoon SC, Choi MH (1999) Local sequence dependence of polyhydroxyalkanoic acid degradation in Hydrogenophaga pseudoflava. J Biol Chem 274:37800–37808

  114. York GM, Lupberger J, Tian J, Lawrence AG, Stubbe J, Sinskey AJ (2003) Ralstonia eutropha H16 encodes two and possibly three intracellular poly[d-(−)-3-hydroxybutyrate] depolymerase genes. J Bacteriol 185:3788–3794

Download references

Acknowledgements

This work was supported by a grant of the Deutsche Forschungsgemeinschaft. I thank M. Unsin and Lei Wang for performing experiments described in Figs. 1 and 2 and W. Armbruster for ESI-MS analysis.

Author information

Correspondence to Dieter Jendrossek.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Jendrossek, D. Peculiarities of PHA granules preparation and PHA depolymerase activity determination. Appl Microbiol Biotechnol 74, 1186–1196 (2007). https://doi.org/10.1007/s00253-007-0860-9

Download citation

Keywords

  • PHB
  • PHA
  • Intracellular PHB depolymerase
  • Native PHB
  • Denatured and artificial PHB
  • PHB depolymerase products