Skip to main content
Log in

Complete Genome Sequence of a Novel Polyhydroxyalkanoate (PHA) Producer, Jeongeupia sp. USM3 (JCM 19920) and Characterization of Its PHA Synthases

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

A novel polyhydroxyalkanoate (PHA)-producing bacterium, Jeongeupia sp. USM3 (JCM 19920) was isolated from the limestone soil at Gua Tempurung, Perak, Malaysia. This is the first report on the complete genome sequence for the genus Jeongeupia. This genome consists of a circular chromosome with a size of 3,788,814 bp and contains 3557 genes. Two PHA synthase (phaC) genes encoding for the key enzyme in the polymerization of PHA monomers and other PHA-associated genes were identified from the genome. Phylogenetic analysis of the PhaC protein sequences has revealed that both PhaC1 and PhaC2 of Jeongeupia sp. USM3 are categorized as Class I PHA synthases with 56% similarity to each other. Both of the PHA synthase genes of this isolate were cloned and heterologously expressed in a PHA mutant strain Cupriavidus necator PHB4. The ability of the transformants to accumulate PHA showed that both PhaC1 and PhaC2 were functional.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Bhubalan K, Yong KH, Kam YC, Sudesh K (2010) Cloning and expression of the PHA synthase gene from a locally isolated Chromobacterium sp. USM2. Malays J Microbiol 6:81–90

    Google Scholar 

  2. Braunegg G, Sonnleitner BY, Lafferty RM (1978) A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass. Eur J Appl Microbiol Biotechnol 6:29–37

    CAS  Google Scholar 

  3. Budde CF, Riedel SL, Willis LB, Rha C, Sinskey AJ (2011) Production of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) from plant oil by engineered Ralstonia eutropha strains. Appl Environ Microbiol 77:2847–2854

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Carpa R, Butiuc-Keul A, Lupan I, Barbu-Tudoran L, Muntean V, Dobrotă C (2012) Poly-β-hydroxybutyrate accumulation in bacterial consortia from different environments. Can J Microbiol 58:660–667

    CAS  PubMed  Google Scholar 

  5. Cevallos MA, Encarnacion S, Leija A, Mora Y, Mora J (1996) Genetic and physiological characterization of a Rhizobium etli mutant strain unable to synthesize poly-beta-hydroxybutyrate. J Bacteriol 178:1646–1654

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Chee JY, Yoga SS, Lau NS, Ling SC, Abed RMM, Sudesh K (2010) Bacterially produced polyhydroxyalkanoate (PHA): converting renewable resources into bioplastics. Curr Res Technol Educ Topics Appl Microbiol Microb Biotechnol 2:1395–1404

    Google Scholar 

  7. Chek MF, Kim SY, Mori T, Arsad H, Samian MR, Sudesh K, Hakoshima T (2017) Structure of polyhydroxyalkanoate (PHA) synthase PhaC from Chromobacterium sp USM2, producing biodegradable plastics. Sci Rep 7:5312

    PubMed  PubMed Central  Google Scholar 

  8. Chen GQ, Wu Q (2005) The application of polyhydroxyalkanoates as tissue engineering materials. Biomaterials 26:6565–6578

    CAS  PubMed  Google Scholar 

  9. Chen WM, Chang RC, Cheng CY, Shiau YW, Sheu SY (2013) Jeongeupia chitinilytica sp. nov., a chitinolytic bacterium isolated from soil. Int J Syst Evol Microbiol 63:934–938

    CAS  PubMed  Google Scholar 

  10. Chin CS, Alexander DH, Marks P, Klammer AA, Drake J, Heiner C, Clum A, Copeland A, Huddleston J, Eichler EE, Turner SW, Korlach J (2013) Nonhybrid, finished microbial genome assemblies from long-read SMRT sequencing data. Nat Methods 10:563

    CAS  Google Scholar 

  11. Demirdöğen RE, Emen FM, Ocakoglu K, Murugan P, Sudesh K, Avşar G (2018) Green nanotechnology for synthesis and characterization of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) nanoparticles for sustained bortezomib release using supercritical CO2 assisted particle formation combined with electrodeposition. Int J Biol Macromol 107:436–445

    PubMed  Google Scholar 

  12. Doi Y, Kitamura S, Abe H (1995) Microbial synthesis and characterization of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate). Macromolecules 28:4822-4828.

    CAS  Google Scholar 

  13. Foong CP, Lau NS, Deguchi S, Toyofuku T, Taylor TD, Sudesh K, Matsui M (2014) Whole genome amplification approach reveals novel polyhydroxyalkanoate synthases (PhaCs) from Japan Trench and Nankai Trough seawater. BMC Microbiol 14:318

    PubMed  PubMed Central  Google Scholar 

  14. Foong CP, Lakshmanan M, Abe H, Taylor TD, Foong SY, Sudesh K (2018) A novel and wide substrate specific polyhydroxyalkanoate (PHA) synthase from unculturable bacteria found in mangrove soil. J Polym Res 25:23

    Google Scholar 

  15. Friedrich B, Hogrefe C, Schlegel HG (1981) Naturally occurring genetic transfer of hydrogen-oxidizing ability between strains of Alcaligenes eutrophus. J Bacteriol 147:198–205

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Hong JW, Song HS, Moon YM, Hong YG, Bhatia SK, Jung HR, Choi TR, Yang SY, Park HY, Choi YK, Yang YH (2019) Polyhydroxybutyrate production in halophilic marine bacteria Vibrio proteolyticus isolated from the Korean peninsula. Bioprocess Biosyst Eng 42:603–610

    CAS  PubMed  Google Scholar 

  17. Kim J, Kim YJ, Choi SY, Lee SY, Kim KJ (2017) Crystal structure of Ralstonia eutropha polyhydroxyalkanoate synthase C-terminal domain and reaction mechanisms. Biotechnol J 12:1600648

    Google Scholar 

  18. Kolibachuk D, Miller A, Dennis D (1999) Cloning, molecular analysis, and sxpression of the polyhydroxyalkanoic acid synthase (phaC) gene from Chromobacterium violaceum. Appl Environ Microbiol 65:3561–3565

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM II, Peterson KM (1995) Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 166:175–176

    CAS  PubMed  Google Scholar 

  20. Kundrat V, Cernekova N, Kovalcik A, Enev V, Marova I (2019) Drug release kinetics of electrospun PHB meshes. Materials 12:1924

    CAS  PubMed Central  Google Scholar 

  21. Lok C (2015) Mining the microbial dark matter. Nat News 522:270

    CAS  Google Scholar 

  22. Maehara A, Ueda S, Nakano H, Yamane T (1999) Analyses of a polyhydroxyalkanoic acid granule-associated 16-kilodalton protein and its putative regulator in the pha locus of Paracoccus denitrificans. J Bacteriol 181:2914–2921

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Mandon K, Michel-Reydellet N, Encarnación S, Kaminski PA, Leija A, Cevallos MA, Elmerich C, Mora J (1998) Poly-β-hydroxybutyrate turnover in Azorhizobium caulinodans is required for growth and affects nifA expression. J Bacteriol 180:5070–5076

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Ng LM, Sudesh K (2016) Identification of a new polyhydroxyalkanoate (PHA) producer Aquitalea sp. USM4 (JCM 19919) and characterization of its PHA synthase. J Biosci Bioeng 122:550–557

    CAS  PubMed  Google Scholar 

  25. Peoples OP, Sinskey AJ (1989) Poly-beta-hydroxybutyrate (PHB) biosynthesis in Alcaligenes eutrophus H16. Identification and characterization of the PHB polymerase gene (phbC). J Biol Chem 264:15298–15303

    CAS  PubMed  Google Scholar 

  26. Pohlmann A, Fricke WF, Reinecke F, Kusian B, Liesegang H, Cramm R, Eitinger T, Ewering C, Pötter M, Schwartz E, Strittmatter A, Voß I, Gottschalk G, Steinbüchel A, Friedrich B, Bowien B (2006) Genome sequence of the bioplastic-producing “Knallgas” bacterium Ralstonia eutropha H16. Nat Biotechnol 24:1257

    PubMed  Google Scholar 

  27. Raberg M, Voigt B, Hecker M, Steinbüchel A (2014) A closer look on the polyhydroxybutyrate-(PHB-) negative phenotype of Ralstonia eutropha PHBˉ4. PLoS ONE ONE 9:e95907

    Google Scholar 

  28. Rehm BHA (2003) Polyester synthases: natural catalysts for plastics. Biochemical Journal 376:15–33

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Rehm BHA, Steinbüchel A (1999) Biochemical and genetic analysis of PHA synthases and other proteins required for PHA synthesis. Int J Biol Macromol 25:3–19

    CAS  PubMed  Google Scholar 

  30. Schlegel HG, Lafferty R, Krauss I (1970) Bacterial mutants of Hydrogenomonas lacking poly-β-hydroxybutyric acid. Experientia 26:554–555

    CAS  PubMed  Google Scholar 

  31. Schlegel HG, Lafferty R, Krauss I (1970) The isolation of mutants not accumulating poly-β-hydroxybutyric acid. Archiv für Mikrobiologie 71:283–294

    CAS  PubMed  Google Scholar 

  32. Schubert P, Steinbüchel A, Schlegel HG (1988) Cloning of the Alcaligenes eutrophus genes for synthesis of poly-beta-hydroxybutyric acid (PHB) and synthesis of PHB in Escherichia coli. J Bacteriol 170:5837–5847

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Shah AA, Hasan F, Hameed A, Ahmed S (2008) Biological degradation of plastics: a comprehensive review. Biotechnol Adv 26:246–265

    CAS  PubMed  Google Scholar 

  34. Simon R, Priefer U, Pühler A (1983) A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Nat Biotechnol 1:784

    CAS  Google Scholar 

  35. Slater SC, Voige WH, Dennis DE (1988) Cloning and expression in Escherichia coli of the Alcaligenes eutrophus H16 poly-beta-hydroxybutyrate biosynthetic pathway. J Bacteriol 170:4431–4436

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Tai YT, Foong CP, Najimudin N, Sudesh K (2016) Discovery of a new polyhydroxyalkanoate synthase from limestone soil through metagenomic approach. J Biosci Bioeng 121:355–364

    CAS  PubMed  Google Scholar 

  37. Tan GYA, Chen CL, Li L, Ge L, Wang L, Razaad IMN, Li Y, Zhao L, Mo Y, Wang JY (2014) Start a research on biopolymer polyhydroxyalkanoate (PHA): a review. Polymers 6:706–754

    Google Scholar 

  38. Tombolini R, Povolo S, Buson A, Squartini A, Nuti MP (1995) Poly-β-hydroxybutyrate (PHB) biosynthetic genes in Rhizobium meliloti 41. Microbiology 141:2553–2559

    CAS  PubMed  Google Scholar 

  39. Valentin HE, Steinbüchel A (1993) Cloning and characterization of the Methylobacterium extorquens polyhydroxyalkanoic-acid-synthase structural gene. Appl Microbiol Biotechnol 39:309–317

    CAS  PubMed  Google Scholar 

  40. Wang Z, Itoh Y, Hosaka Y, Kobayashi I, Nakano Y, Maeda I, Umeda F, Yamakawa J, Kawase M, Yag K (2003) Novel transdermal drug delivery system with polyhydroxyalkanoate and starburst polyamidoamine dendrimer. J Biosci Bioeng 95:541–543

    CAS  PubMed  Google Scholar 

  41. Wei DX, Dao JW, Chen GQ (2018) A micro-ark for cells: highly open porous polyhydroxyalkanoate icrospheres as injectable scaffolds for tissue regeneration. Adv Mater 30:1802273

    Google Scholar 

  42. Wittenborn EC, Jost M, Wei Y, Stubbe J, Drennan CL (2016) Structure of the catalytic domain of the class I polyhydroxybutyrate synthase from Cupriavidus necator. J Biol Chem 291:25264–25277

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Yoon JH, Choi JH, Kang SJ, Choi NS, Lee JS, Song JJ (2010) Jeongeupia naejangsanensis gen. nov., sp. nov., a cellulose-degrading bacterium isolated from forest soil from Naejang Mountain in Korea. Int J Syst Evol Microbiol 60:615–619

    CAS  PubMed  Google Scholar 

  44. Zinn M, Witholt B, Egli T (2001) Occurrence, synthesis and medical application of bacterial polyhydroxyalkanoate. Adv Drug Deliv Rev 53:5–21

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by USM-RIKEN International Centre for Aging Science (URICAS) (1001/PBIOLOGI/870033). N.-A.A.Z. is grateful to the Public Service Department of Malaysia scholarship scheme. Authors thank Hui Pheng Kho for her contribution in experimental works and Dr. Todd Taylor (RIKEN, Japan) for his useful advice.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kumar Sudesh.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 207 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zain, NA.A., Ng, LM., Foong, C.P. et al. Complete Genome Sequence of a Novel Polyhydroxyalkanoate (PHA) Producer, Jeongeupia sp. USM3 (JCM 19920) and Characterization of Its PHA Synthases. Curr Microbiol 77, 500–508 (2020). https://doi.org/10.1007/s00284-019-01852-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00284-019-01852-z

Navigation