Isolation and characterization of polyhydroxyalkanoate producing bacteria from legume rhizosphere soils in Me Linh commune, Hanoi

Kim Thoa Nguyen, Thi Hong Nhung Lai, Thi Tuyet Minh Phan, Thi Minh Tu Hoa, Thanh Thuy Tran, Quoc Viet Nguyen, The Trang Nguyen, Thi Thanh Trung Do, Tat Thanh Le
Author affiliations

Authors

  • Kim Thoa Nguyen \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
    \(^2\)Graduate University of Science and Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietna
    https://orcid.org/0000-0002-2702-1907
  • Thi Hong Nhung Lai \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Thi Tuyet Minh Phan \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Thi Minh Tu Hoa \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Thanh Thuy Tran \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Quoc Viet Nguyen \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • The Trang Nguyen \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam https://orcid.org/0009-0003-2764-122X
  • Thi Thanh Trung Do \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam
  • Tat Thanh Le \(^1\) Institute of Biology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Hanoi, Vietnam

DOI:

https://doi.org/10.15625/vjbt-23144

Keywords:

Bioplastic, polyhydroxyalkanoates (PHAs), Priestia spp., Pseudomonas spp., soil bacteria

Abstract

The increasing demand for sustainable bioplastics has driven the search for polyhydroxyalkanoate (PHA)-producing microorganisms from diverse ecological niches. In this study, we isolated and characterized native soil bacteria with the ability to accumulate PHAs from the rhizosphere of leguminous crops in Me Linh commune, Hanoi. A total of 206 bacterial isolates were obtained from eight soil samples and screened for intracellular PHA granules using Sudan Black B and Nile Blue A staining. Of these, 21 isolates tested positive for PHA accumulation, and five strains (ML53, ML71, ML91, ML113, and ML205) were selected for further analysis. Quantitative fermentation experiments revealed that strains ML113, ML91, and ML71 achieved the highest PHA contents, ranging from 20.7% to 24.8% of dry cell weight (DCW). Fourier-transform infrared (FTIR) spectroscopy confirmed the presence of functional groups characteristic of PHB and PHBV biopolymers. Morphological, biochemical, and molecular analyses—particularly 16S rRNA gene sequencing and phylogenetic tree construction—revealed that the selected isolates belong to three distinct genera: Pseudomonas (ML53, closely related to P. putida), Micrococcus (ML91, closely related to M. luteus), and Priestia (ML71 and ML205, clustered with P. megaterium, and ML113, closely related to P. aryabhattai). The results highlight the legume rhizosphere as a rich source of genetically diverse and metabolically competent PHA-producing bacteria with promising potential for future bioplastic development.

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References

Arcos-Hernández, M. V., Laycock, B., Donose, B. C., Pratt, S., Halley, P., Al-Luaibi, S., et al. (2013). Physicochemical and mechanical properties of mixed culture polyhydroxyalkanoate (PHBV). European Polymer Journal, 49(4), 904-913. https://doi.org/10.1016/j.eurpolymj.2012.10.025

Barillot, C. D. C., Sarde, C. O., Bert, V., Tarnaud, E., & Cochet, N. (2013). A standardized method for the sampling of rhizosphere and rhizoplan soil bacteria associated to a herbaceous root system. Annals of Microbiology, 63, 471–476. https://doi.org/10.1007/s13213-012-0491-y

Belal, E. B. (2013). Production of poly-β-hydroxybutyric acid (PHB) by Rhizobium elti and Pseudomonas stutzeri. Current Research Journal of Biological Sciences, 5(6), 273-284. http://dx.doi.org/10.19026/crjbs.5.5429

Chen, G. Q. (2010). Plastics completely synthesized by bacteria: Polyhydroxyalkanoates. In Plastics from Bacteria, 17–37. https://doi.org/10.1007/978-3-642-03287-5_2

Chen, J. Y., Song, G. & Chen, G. Q. (2006). A lower specificity PhaC2 synthase from Pseudomonas stutzeri catalyses the production of copolyesters consisting of short-chain-length and medium-chain-length 3-hydroxyalkanoates. Antonie Van Leeuwenhoek, 89, 157-167. https://doi.org/10.1007/s10482-005-9019-9

Claus, D., & Berkeley, R. C. W. (1986). Genus Bacillus Cohn 1872, in: Bergey’s Manual of Systematic Bacteriology, Vol. 2 (P. H. A. Sneath, ed.), Bergey’s Manual Trust, Williams and Wilkins, Baltimore, 1105–1139.

Getino, L., Martín, J. L., & Chamizo-Ampudia, A. (2024). A review of polyhydroxyalkanoates: Characterization, production, and application from waste. Microorganisms, 12(10), 2028. https://doi.org/10.3390/microorganisms12102028

Israni, N., & Shivakumar, S. (2015). Evaluation of Upstream Process parameters influencing the growth associated PHA accumulation in Bacillus sp. Ti3. Journal of Scientific & Industrial Research, 74, 290-295.

Joyline, M., & Aruna, K. (2019). Production and characterization of polyhydroxyalkanoates (PHA) by Bacillus megaterium strain JHA using inexpensive agro-industrial wastes. International Journal of Scientific Research, 10, 33359–33374. http://dx.doi.org/10.24327/ijrsr.2019.1007.3656

Koller, M., & Rodríguez-Contreras, A. (2015). Techniques for tracing PHA-producing organisms and for qualitative and quantitative analysis of intra- and extracellular PHA. Engineering in Life Sciences, 15, 558-581. https://doi.org/10.1002/elsc.201400228

Laycock, B., Halley, P., Pratt, S., Werker, A., & Lant, P. (2013). The chemomechanical properties of microbial polyhydroxyalkanoates. Progress in Polymer Science, 38(3–4), 536–583. https://doi.org/10.1016/j.progpolymsci.2012.06.003

Lee, S. Y. (1996). Bacterial polyhydroxyalkanoates. Biotechnology and Bioengineering, 49(1), 1–14. https://doi.org/10.1002/(SICI)1097-0290(19960105)49:1%3C1::AID-BIT1%3E3.0.CO;2-P

Li, M. L., Doudin, K., Robins, D. B., Tetradis-Mairis, G., Wong, T. S., Tee, K. L. (2025). Microbial synthesis of polyhydroxyalkanoate blends with engineered Pseudomonas putida, New Biotechnology, 88, 161-170. https://doi.org/10.1016/j.nbt.2025.05.004

Madigan, M. T., Bender, K. S., Buckley, D. H., Sattley, W. M., & Stahl, D. A. (2018). Brock Biology of Microorganisms (15th ed.). Pearson.

Musa, H., Bolanle, B. B., Kasim, F. H., Arbain, D. (2016). Screening and production of Polyhydroxybutyrate (PHB) by bacterial strains isolated from rhizosphere soil of groundnut plants. Sains Malaysiana, 45, 1469-1476.

Nishida, M., Tanaka, T., Hayakawa, Y., Nishida, M. (2018). Solid state nuclear magnetic resonance (NMR) and nuclear magnetic relaxation time analyses of molecular mobility and compatibility of plasticized polyhydroxyalkanoates (PHA) copolymers. Polymers, 10(5), 506. https://doi.org/10.3390/polym10050506

Orita, I., Iwazawa, R., Nakamura, S., & Fukui, T. (2012). Identification of mutation points in Cupriavidus necator NCIMB 11599 and genetic reconstitution of glucose-utilization ability in wild strain H16 for polyhydroxyalkanoate production. Journal of Bioscience and Bioengineering, 113(1), 63–69. https://doi.org/10.1016/j.jbiosc.2011.09.014

Ostle, A. G., Holt, J. G. (1982). Nile blue A as a fluorescent stain for poly-beta-hydroxybutyrate. Applied and Environmental Microbiology, 44(1), 238-241. https://journals.asm.org/doi/10.1128/aem.44.1.238-241.1982

Palleroni, N. J. (2010). The Pseudomonas story. Environmental Microbiology, 12(6), 1377–1383. https://doi.org/10.1111/j.1462-2920.2009.02041.x

Patil, T. D., Ghosh, S., Agarwal, A., Patel, S. K. S., Tripathi, A. D., Mahato, D. K., et al. (2024). Production, optimization, scale up and characterization of polyhydoxyalkanoates copolymers utilizing dairy processing waste. Scientific Reports, 14(1), 1620. https://doi.org/10.1038/s41598-024-52098-0

Râpă, M., Stefan, L. M., Seciu-Grama, A.-M., Gaspar-Pintiliescu, A., Matei, E., Zaharia, C., et al. (2022). Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(3HB-co-3HV))/Bacterial Cellulose (BC) biocomposites for potential use in biomedical applications. Polymers, 14(24), 5544. https://doi.org/10.3390/polym14245544

Rivera-Briso A. L. & Serrano-Aroca Á. (2018). Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate) Enhancement strategies for advanced applications. Polymers (Basel), 10(7), 732. https://doi.org/10.3390/polym10070732

Schlegel, H. G., Lafferty, R., & Krauss, I. (1970). The isolation of mutants not accumulating poly-β-hydroxybutyric acid. Archiv für Mikrobiologie, 71(3), 283–294. https://doi.org/10.1007/BF00410161

Shah, S., & Kumar, A. (2021). Production and characterization of polyhydroxyalkanoates from industrial waste using soil bacterial isolates. Brazilian Journal of Microbiology, 52(2), 715–726. https://doi.org/10.1007/s42770-021-00452-z

Spiekermann, P., Rehm, B. H., Kalscheuer, R., Baumeister, D., & Steinbüchel, A. (1999). A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds. Archives of Microbiology, 171(2), 73–80. https://doi.org/10.1007/s002030050681

Trakunjae, C., Sudesh, K., Neoh, S. Z., Boondaeng, A., Apiwatanapiwat, W., Janchai, P., et al. (2022). Biosynthesis of P(3HB-co-3HHx) copolymers by a newly engineered strain of Cupriavidus necator PHB−4/pBBR_CnPro-phaCRp for skin tissue engineering application. Polymers, 14(19), 4074. https://doi.org/10.3390/polym14194074

Zhang, J., & Cran, M. J. J. (2022). Production of polyhydroxyalkanoate nanoparticles using a green solvent. Applied Polymer Science, 139(23), e52319. https://doi.org/10.1002/app.52319

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Published

30-09-2025

How to Cite

Nguyen, K. T., Lai, T. H. N., Phan, T. T. M., Hoa, T. M. T., Tran, T. T., Nguyen, Q. V., … Le, T. T. (2025). Isolation and characterization of polyhydroxyalkanoate producing bacteria from legume rhizosphere soils in Me Linh commune, Hanoi. Vietnam Journal of Biotechnology, 23(3), 415–428. https://doi.org/10.15625/vjbt-23144

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