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Bacillus cereus saba.zh, a novel bacterial strain for the production of bioplastic (polyhydroxybutyrate)

  • Environmental Microbiology - Research Paper
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Abstract

The identification of novel bacterial strains with a high production potential of polyhydroxybutyrate (PHB) to substitute the bioplastics with non-biodegradable plastics and reducing environmental pollution is really effective. The present study was done with the purpose of PHB bioplastic production using a novel bacterial strain. Twenty-one different bacterial isolates were obtained from petrochemical wastewater, which among them, 10 isolates were PHB positive. The presence of PHB granules was detected in the isolates using Sudan Black B staining. The most excellent PHB-accumulating bacterium with a maximum yield of PHB (61.53%) was selected and identified as Bacillus cereus saba.zh, based on morphological, biochemical, and molecular techniques. 16S rRNA nucleotide sequence of the bacterium was assigned accession number: MT975245 in the NCBI database. The phylogenetic tree data showed that the closest type strain to the Bacillus cereus saba.zh is the Bacillus cereus SDB4 (91%). The three genes (phaA, phaB, and phaC) responsible for the PHB biosynthesis were amplified using the specific oligonucleotide primers by PCR technique. The highest PHB yield was achieved when the culture medium was supplemented with 3% sugarcane molasses as a carbon source, ammonium sulfate as the nitrogen source, at pH 7, and temperature of 30 °C. The characterization of the extracted polymer by FTIR and 1H NMR spectroscopy proves the presence of methyl, methylene, methine, hydroxyl, and ester carbonyl groups and confirmed the structure of biopolymer as PHB. The novel strain Bacillus cereus saba.zh has good potential as an appropriate candidate for low-cost industrial production of bioplastic.

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References

  1. Urbanek AK, Rymowicz W, Mirończuk AM (2018) Degradation of plastics and plastic-degrading bacteria in cold marine habitats. Appl Microbiol Biotechnol 102:7669–7678. https://doi.org/10.1007/s00253-018-9195-y

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Selvamurugan M, Sivakumar P (2019) Bioplastics - an eco-friendly alternative to petrochemical plastics. Curr World Environ 14:49–59. https://doi.org/10.12944/CWE.14.1.07

    Article  Google Scholar 

  3. Sharma N (2019) Polyhydroxybutyrate (PHB) production by bacteria and its application as biodegradable plastic in various industries. Academ J Polym Sci 2:001–003. https://doi.org/10.19080/AJOP.2019.02.555587

    Article  CAS  Google Scholar 

  4. Chandegara VK, Cholera SP, Nandasana JN, Kumpavat MT, Patel KC (2015) Plastic packaging waste impact on climate change and its mitigation. In: Subbaiah R, Prajapati GV (eds) Water management and climate smart agriculture. Adaptation of climatic resilient water management and agriculture.Gyan Publishing House, New Delhi, pp 404–415

    Google Scholar 

  5. Yu J, Chen LXL (2008) The greenhouse gas emissions and fossil energy requirement of bioplastics from cradle to gate of a biomass refinery. Environ Sci Technol 42:6961–6966. https://doi.org/10.1021/es7032235

    Article  CAS  PubMed  Google Scholar 

  6. Muhammadi S, Afzal M, Hameed S (2015) Bacterial polyhydroxyalkanoates-eco-friendly next generation plastic: production, biocompatibility, biodegradation, physical properties and applications. Green Chem Lett Rev 8:56–77. https://doi.org/10.1080/17518253.2015.1109715

    Article  CAS  Google Scholar 

  7. Lathwal P, Nehra K, Singh M, Jamdagni P, Raba JS (2015) Optimization of culture parameters for maximum polyhydroxybutyrate production by selected bacterial strains isolated from rhizospheric soils. Polish J Microbiol 64:227–239. https://doi.org/10.5604/01.3001.0009.2118

    Article  Google Scholar 

  8. Nehra K, Jaglan A, Shaheen A, Yadav J, Lathwal P (2015) Production of poly-β- hydroxybutyrate (PHB) by bacteria isolated from rhizospheric soils. Int J Microbial Res Technol 2:38–48

    Google Scholar 

  9. Kalia VC, Ray S, Patel SK, Singh M, Singh GP (2019) Biotechnological applications of polyhydroxyalkanoates: applications of polyhydroxyalkanoates and their metabolites as drug carriers. Springer, Singapore, pp 35–48. https://doi.org/10.1007/978-981-13-3759-8

    Book  Google Scholar 

  10. Maheswari MU, Dhandayuthapani K (2013) Fermentation opimization for production of polyhydroxybutyrate (PHB) by newly isolated Azotobacter vinelandii KDP. Int J Pharm Bio Sci 4:779–787

    Google Scholar 

  11. Mahitha G, Madhuri R (2015) Purification and characterization of polyhydroxybutyrate produced from marine bacteria. Int J Sci Eng Res 6:71–75

    Google Scholar 

  12. Thammasittirong A, Saechow S, Thammasittirong SNR (2017) Efficient polyhydroxybutyrate production from Bacillus thuringiensis using sugarcane juice substrate. Turk J Biol 41:992–1002. https://doi.org/10.3906/biy-1704-13

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Mostafa YS, Alrumman SA, Otaif KHD, Alamri SA, Mostafa MS, Sahlabji T (2020) Production and characterization of bioplastic by polyhydroxybutyrate accumulating Erythrobacter aquimaris isolated from mangrove rhizosphere. Molecules 25:1–20. https://doi.org/10.3390/molecules25010179

    Article  CAS  Google Scholar 

  14. Chandani N, Mazumder PB, Bhattacharjee A (2018) Biosynthesis of biodegradable polymer by a potent soil bacterium from a stress-prone environment. J App Biol Biotech 6:54–60. https://doi.org/10.7324/JABB.2018.60209

    Article  CAS  Google Scholar 

  15. Doetsch RN (1981) Determinative methods of light microscopy. In: Gerhardt P, Murray RGE, Costilow RN, Nester EW, Wood WA, Krieg NR, Phillips GB (eds) Manual of methods for general bacteriology, 2nd edn. ASM Press, Washington, pp 22–31

    Google Scholar 

  16. Bergey DH, Holt JG (1994) Bergey’s manual of determinative bacteriology. Williams & Wilkins, Baltimore

    Google Scholar 

  17. Marin M, Garcia-Lechuz JM, Alonso P, Villanueva M, Alcalá L, Gimeno M, Cercenado E, Sánchez- Somolinos M, Radice C, Bouza E (2012) Role of universal 16S rRNA gene PCR and sequencing in diagnosis of prosthetic joint infection. J Clin Microbiol 50:583–589. https://doi.org/10.1128/JCM.00170-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Hassan MA, Bakhiet EK, Ali SG, Hussien HR (2016) Production and characterization of polyhydroxybutyrate (PHB) produced by Bacillus sp. isolated from Egypt. J App Pharm Sci 6:046–051. https://doi.org/10.7324/JAPS.2016.60406

    Article  CAS  Google Scholar 

  19. Mascarenhas J, Aruna K (2017) Screening of polyhydroxyalkonates (PHA) accumulating bacteria from diverse habitats. J global biosci 6:4835–4848

    Google Scholar 

  20. Sharmila M, Durgesh DW, Tumane PM (2017) Optimization and characterization of bioplastic produced by Bacillus Cereus. Int J Recent Sci Res 8:16565–16571. https://doi.org/10.24327/ijrsr.2017.0804.0173

    Article  Google Scholar 

  21. Valappil SP, Boccaccini AR, Bucke C, Roy I (2007) Polyhydroxyalkanoates in gram- positive bacteria: insights from the genera Bacillus and Streptomyces. Antonie Van Leeuwenhoek 91:1–17. https://doi.org/10.1007/s10482-006-9095-5

    Article  CAS  PubMed  Google Scholar 

  22. Moorkoth D, Nampoothiri KM (2016) Production and characterization of poly (3-hydroxy butyrate-co-3 hydroxyvalerate) (PHBV) by a novel halotolerant mangrove isolate. Bioresource Technol 201:253–260. https://doi.org/10.1016/j.biortech.2015.11.046

    Article  CAS  Google Scholar 

  23. Clarridge JE (2004) Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev 17:840–862. https://doi.org/10.1128/CMR.17.4.840-862.2004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Mayeli N, Motamedi H, Heidarizadeh F (2015) Production of polyhydroxybutyrate by Bacillus axaraqunsis BIPC01 using petrochemical wastewater as carbon source. Braz Arch Biol Technol 58:643–650. https://doi.org/10.1590/S1516-8913201500048

    Article  CAS  Google Scholar 

  25. Aarthi N, Ramana KV (2011) Identification and characterization of polyhydroxybutyrate producing Bacillus cereus and Bacillus mycoides strains. Int J Env Sci 1:744–756

    CAS  Google Scholar 

  26. Sathiyanarayanan G, Kiran GS, Selvin J, Saibaba G (2013) Optimization of polyhydroxybutyrate production by marine Bacillus megaterium MSBN04 under solid state culture. Int J Biol Macromol 60:253–261. https://doi.org/10.1016/j.ijbiomac.2013.05.031

    Article  CAS  PubMed  Google Scholar 

  27. Sathiyanarayanan G, Saibaba G, Kiran GS, Selvin J (2013) Process optimization and production of polyhydroxybutyrate using palm jaggery as economical carbon source by marine sponge-associated Bacillus licheniformis MSBN12. Bioprocess Biosyst Eng 36:1817–1827. https://doi.org/10.1007/s00449-013-0956-9

    Article  CAS  PubMed  Google Scholar 

  28. Poonam RB, Shruti UP, Kishor PU, Basavaraj SH (2015) Production and characterization of thermostable polyhydroxybutyrate from Bacillus cereus PW3A. J Biochem Tech 6:990–995

    Google Scholar 

  29. Korotkova N, Lidstrom ME (2001) Connection between poly-β-hydroxybutyrate biosynthesis and growth on C1 and C2 compounds in the methylotroph Methylobacterium extorquens AM1. J Bacteriol 183:1038–1046. https://doi.org/10.1128/JB.183.3.1038-1046.2001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Alarfaj AA, Arshad M, Sholkamy EN, Munusamy MA (2015) Extraction and characterization of polyhydroxybutyrates (PHB) from Bacillus thuringiensis KSADL127 isolated from mangrove environments of Saudi Arabia. Braz Arch Biol Technol 58:781–788. https://doi.org/10.1590/S1516-891320150500003

    Article  CAS  Google Scholar 

  31. Berekaa MM, Thawadi AMA (2012) Biosynthesis of polyhydroxybutyrate (PHB) biopolymer by Bacillus megaterium SW1-2: application of Box-Behnken design for optimization of process parameters. Afr J Microbiol Res 6:838–845. https://doi.org/10.5897/AJMR-11-1382

    Article  CAS  Google Scholar 

  32. Das R, Dey A, Pal A, Paul AK (2016) Influence of growth conditions on production of poly (3-hydroxybutyrate) by Bacillus cereus HAL 03 endophytic to Helianthus annuus L. J App Biol Biotech 4:075–084. https://doi.org/10.7324/JABB.2016.40409

    Article  CAS  Google Scholar 

  33. Nabila FS, VeenaGayathri K (2017) Isolation and screening of Phb producing halotolerant bacterial strains from a saline environment. J Biotech Biomed Sci 1:34–45. https://doi.org/10.14302/ISSN.2576-6694.JBBS-17-1783

    Article  Google Scholar 

  34. Bhuwal AK, Singh G, Aggarwal NK, Goyal V, Yadav A (2014) Poly-β-hydroxybutyrate production and management of cardboard industry effluent by new Bacillus sp. NA10. Biores Bioprocess 1:1–11. https://doi.org/10.1186/s40643-014-0009-5

    Article  Google Scholar 

  35. Mohandas SP, Balan L, Lekshmi N, Cubelio SS, Philip R, Bright Singh IS (2016) Production and characterization of polyhydroxybutyrate from Vibrio harveyi MCCB 284 utilizing glycerol as carbon source. J App Microbiol 122:698–707. https://doi.org/10.1111/jam.13359

    Article  CAS  Google Scholar 

  36. Devi AB, Nachiyar CV, Kaviyarasi T, Samrot AV (2015) Characterization of polyhydroxybutyrate synthesized by Bacillus cereus. Int J Pharm Pharm Sci 7:140–144

    CAS  Google Scholar 

  37. Sathiyanarayanan G, Saibaba G, Kiran GS, Selvin J (2013) A statistical approach for optimization of polyhydroxybutyrate production by marine Bacillus subtilis MSBN17. Int J Biol Macromol 59:170–177. https://doi.org/10.1016/j.ijbiomac.2013.04.040

    Article  CAS  PubMed  Google Scholar 

  38. Oliveira F, Freire D, Castilho L (2004) Production of poly (3-hydroxybutyrate) by solid-state fermentation with Ralstonia eutropha. Biotechnol Lett 26:1851–1855. https://doi.org/10.1007/s10529-004-5315-0

    Article  CAS  PubMed  Google Scholar 

  39. Teclu D, Tivchev G, Laing M, Wallis M (2008) Determination of the elemental composition of molasses and its suitability as carbon source for growth of sulphate-reducing bacteria. J Hazard Mater 161:1157–1165. https://doi.org/10.1016/j.jhazmat.2008.04.120

    Article  CAS  PubMed  Google Scholar 

  40. Oderinde R, Ngoka L, Adesogan K (1986) Comparative study of the effect of ferrocyanide and EDTA on the production of ethyl alcohol from molasses by Saccharomyces cerevisiae. Biotechnol Bioeng 28:1156–1162. https://doi.org/10.1002/bit.260281003

    Article  Google Scholar 

  41. El-Sheekh MM, El-Abd MA, El-Diwany AI, Abdel-mohsen S, Ismail AS, Omar TH (2015) Poly-3-hydroxybutyrate (PHB) production by Bacillus flexus ME-77 using some industrial wastes. Rend Fis Acc Lincei 26:109–119. https://doi.org/10.1007/s12210-014-0368-z

    Article  Google Scholar 

  42. Gabr GA (2018) Isolation and identification of bacterial strains able to biopolymer polyhydroxybutyrate (Phb) production from soil of Al-Kharj probes, Saudi Arabia. J Pharm Res Int 21:1–11. https://doi.org/10.9734/JPRI/2018/39532

    Article  Google Scholar 

  43. Mostafa YS, Alrumman SA, Alamri SA, Otaif KA, Mohamed S, Mostafa MS, Alfaify AM (2020) Bioplastic (poly-3-hydroxybutyrate) production by the marine bacterium Pseudodonghicola xiamenensis through date syrup valorization and structural. Nature 10:1–13. https://doi.org/10.1038/s41598-020-65858-5

    Article  CAS  Google Scholar 

  44. Getachew A, Woldesenbet F (2016) Production of biodegradable plastic by polyhydroxybutyrate (PHB) accumulating bacteria using low cost agricultural waste material. BMC Res Notes 9:1–9. https://doi.org/10.1186/s13104-016-2321-y

    Article  CAS  Google Scholar 

  45. Rehman A, Aslam A, Masood R, Aftab MN, Ajmal R, Haq IU (2016) Production and characterization of a thermostable bioplastic (poly-β- hydroxybutyrate) from Bacillus cereus NRRL-B-3711. Pak J Bot 48:349–356

    Google Scholar 

  46. Alshehrei F (2019) Production of polyhydroxybutyrate (PHB) by bacteria isolated from soil of Saudi Arabia. J Pure Appl Microbiol 13:897–904. https://doi.org/10.22207/JPAM.13.2.26

    Article  Google Scholar 

  47. Al-Battashi H, Annamalai N, Al-Kindi Sh, Nair AS, Al-Bahry S, Verma JP, Sivakumar N (2019) Production of bioplastic (poly-3-hydroxybutyrate) using waste paper as a feedstock: optimization of enzymatic hydrolysis and fermentation employing Burkholderia sacchari. J clean Product 214:236–247. https://doi.org/10.1016/j.jclepro.2019.12.239

    Article  CAS  Google Scholar 

  48. Elsayed BB (2013) Production of poly-β-hydroxybutyric acid (PHB) by Rhizobium elti and Pseudomonas stutzeri. Curr Res J Biol Sci 5:273–284. https://doi.org/10.19026/crjbs.5.5429

    Article  Google Scholar 

  49. Aly MM, Albureikan MO, Al-robay H, Kabli S (2013) Effects of culture conditions on growth and poly-β-hydroxybutyric acid production by Bacillus cereus MM7 isolated from soil samples from Saudi Arabia. Life Sci J 10:1884–1891

    Google Scholar 

  50. Saleem F, Aslam R, Saleem Y, Naz S, Syed Q, Munir N, Khurshid N, Shakoori AR (2014) Analysis and evaluation of growth parameters for optimum production of polyhydroxybutyrate (PHB) by Bacillus thuringiensis strain CMBL-BT-6. Pak J Zool 46:1337–1344

    CAS  Google Scholar 

  51. Chandani N, Mazumder PB, Bhattacharjee A (2014) Production of polyhydroxybutyrate (biopolymer) by Bacillus tequilensis NCS-3 isolated from municipal waste areas of Silchar, Assam. Int J Sci Res 3:198–203

    Google Scholar 

  52. Belal EB, Farid MA (2016) Production of poly-β-hydroxybutyric acid (PHB) by Bacillus cereus. Int J curr Microbial App Sci 5:442–460. https://doi.org/10.20546/ijcmas

    Article  CAS  Google Scholar 

  53. Hawas JM, El-Banna T, Belal EBA, El-Azizv AA (2016) Production of bioplastic some selected bacterial strains. Int J Curr Microbiol App Sci 5:10–22. https://doi.org/10.20546/ijcmas

    Article  CAS  Google Scholar 

  54. Hamieh A, Olama Z, Holail H (2013) Microbial production of polyhydroxybutyrate, a biodegradable plastic using agro-industrial waste products. Glo Adv Res J Microbiol 2:54–64

    Google Scholar 

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Acknowledgements

The authors of this essay thank dear personnel of the microbiology research laboratory in Islamic Azad University, Tonekabon Branch, because of their executive supports.

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Authors and Affiliations

  1. Department of Microbiology, Tonekabon Branch, Islamic Azad University, Tonekabon, Iran

    Saba Amiri

  2. Department of Chemistry, Tonekabon Branch, Islamic Azad University, Tonekabon, Iran

    Masoud Mohammadi Zeydi

  3. Department of Biochemistry, Babol Branch, Islamic Azad University, Babol, Iran

    Nasim Amiri

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  1. Saba Amiri
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Amiri, S., Mohammadi Zeydi, M. & Amiri, N. Bacillus cereus saba.zh, a novel bacterial strain for the production of bioplastic (polyhydroxybutyrate). Braz J Microbiol 52, 2117–2128 (2021). https://doi.org/10.1007/s42770-021-00599-9

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