⚠ Official Notice: www.ijisrt.com is the official website of the International Journal of Innovative Science and Research Technology (IJISRT) Journal for research paper submission and publication. Please beware of fake or duplicate websites using the IJISRT name.



Isolation and Identification of Agrobacterium and Azotobacter Species Capable of Producing Curdlan and Alginate Biopolymers


Authors : Mbahi, M. A.; Doughari, J. H.

Volume/Issue : Volume 11 - 2026, Issue 7 - July


Google Scholar : https://tinyurl.com/3w3h45kj

Scribd : https://tinyurl.com/yc3na6ah

DOI : https://doi.org/10.38124/ijisrt/26jul1379

Note : A published paper may take 4-5 working days from the publication date to appear in PlumX Metrics, Semantic Scholar, and ResearchGate.


Abstract : Curdlan is an important homopolysaccharide, while Alginate is a naturally occurring polymer. Numerous bacteria species have the potential to produce these biopolymers.In this study, Agrobacterium and Azotobacter species capable of producing these polymers were isolated and identified. Soil samples from different legume and cereal farms were inoculated unto mannitol glutamate agar containing aniline blue dyes, Nitrogen free Jensen’s agar medium and Asby agar medium for isolation of the bacteria,The isolates obtained were subjected to biochemical tests and 16S rRNA sequencing for identification. Result showed that 5 species of Agrobaterium grew on aneline blue mannitol glutamate agar (aMGA) exhibiting blue coloration; 2 Azotobacter spp grew on Jensen nitrogen free agar; Agrobaterium and Azotobacter species appeared as Gram negative rods; Agrobaterium spp were KOH sensitive, urease, catalase,citrate positive, hydrolyze starch while Azotobacter spp were positive for methly red, indole, catalase, urease, citrate utilization, starch hydrolysis, and hydrogen sulfide production. Molecular identification showed a band of 1500bp product of 16S rRNA gene of Agrobacterium spp on gel electrophoresis. Result further showed that the Agrobacterium species isolates were capable of producing Curdlan while Azotobacter vinelandii demonstrated Alginate biopolymer production. these findings provide suitable indigenous bacterial strains that will enhance biopolymer production.

Keywords : Agrobacterium, Alginate, Azotobacter, Biopolymers, Curdlan.

References :

  1. Al-Rumaidh, Y.S. S., Al-Sahlany, S.T. G. & Ali, H.I. (2023). The Impact of Using Date Juice as a Carbon Source on Curdlan Produced by a Local Isolate of Agrobacterium leguminum. Basrah Journal of Agricultural Sciences, 36(1), 149-163. https://doi.org/10.37077/25200860.2023.36.1.13
  2. Aquinas, N., Bhat, R. & Selvaraj, S. (2021). A review presenting production, characterization, and applications of biopolymer curdlan in food and pharmaceutical sectors. Polymer Bulletin https://doi.org/10.1007/s00289-021-03860-1
  3. Aquinas, N., Bhat, R. M. & Selvaraj, S. (2024). Submerged Fermentation and Kinetics of Newly Isolated Priestia megaterium for the Production of Biopolymer Curdlan. Journal of Polymers and the Environment, 32 (9), 4683–4698. https://doi.org/10.1007/s10924-024-03224-6
  4. Bouzenad, N., Ammouchi, N., Abdennouri, A., Bouzana, A., Rudayni, H.A., Boufahja, F. & Bendif, H. (2025). Comprehensive review of alginate: Sources, synthesis, and application in the food packaging sector. Italian Journal of Food Science, 37: 4, 1–15. https://doi.org/10.15586/ijfs.v37i4.3033
  5. Ferdous, M. L., Hossain, M. N., Ali, M. O., Islam, M. S. & Yasmin, S. (2021). Morphological, biochemical and molecular identification of the wild strain of Agrobacterium tumefaciens from crown gall infected mango tree. Fundamental and Applied Agriculture, 6(1), 43-49. https://doi.org/10.5455/faa.136134
  6. Garcia-Vaquero, M., Rajauria, G., O’Doherty, J.V. & Sweeney, T. (2017). Polysaccharides from macroalgae: Recent advances, innovative technologies and challenges in extraction and purification. Food Research International, Microalgae and Seaweeds as Potential Source of Valuable Nutrients, Food Additives and Nutraceuticals for Human and Animal Consumption, 99: 1011– 1020. https://doi.org/10.1016/j.foodres.2016.11.016
  7. Hurtado, A., Aljabali, A.A.A., Mishra, V., Tambuwala, M.M. and Serrano-Aroca, Á. (2022). Alginate: Enhancement Strategies for Advanced Applications. Int J Mol Sci, 23: 4486. https://doi.org/10.3390/ijms23094486
  8. Ijah, U.J. & Antai, S. P. (2003). The potential use of Chicken drop microorganisms for oil spill remediation.  Environmentalist. 23 (1):89-95.
  9. Jimenez, J.D., Montana, J.S. & Martínez, M.M. (2011). Characterization of free nitrogen fixing bacteria of the genus Azotobacter in organic vegetable-grown Colombian soils. Braz. J. Microbiol. 42(3) 66-78.
  10. Koffi, D., Yapi, G.Y., Kouadjo, K.F., Tossea, S.K., Toure, O.A. & Djaman, J.A. (2023). Implementation of in-house Methods for Isolating Fungal DNA of Clinical Samples. American Journal of Biomedical Research, 11(1):7- 13.
  11. Kouadjo, K.F., Koffi, D., Yaye, Y.G., Kouadio, K.J., Ira-Bonouman, A.V.B., Ouattara, Y.K., Biale, M.D., Alcazar-Fuoli, L., Djaman, A.J. & Alastruey-Izquierdo, A. (2025). Fluconazole susceptibility testing and detection of mutations in the Erg11 gene of Candida albicans clinical isolates in Côte d’Ivoire. African Journal of Microbiology Research, 19 (7), pp. 160-170. https://doi.org/10.5897/AJMR2025.9798
  12. Laxmi, V.M., Latha, D. & Jayasree, A.S. (2018). Production and characterization of Curdlan from Agrobacterium sp. Int. J. Pharm. Sci. & Res., 9(11): 4871-74. https://doi.org/10.13040/IJPSR.0975-8232.9(11).4871-74.
  13. Lee PY, Costumbrado J, Hsu CY, Kim YH. Agarose gel electrophoresis for the separation of DNA
  14. fragments. J Vis Exp. 2012 Apr 20;(62):3923. doi: 10.3791/3923. PMID: 22546956;
  15. PMCID: PMC4846332.
  16. Moselhy, M. A., Lotfy, F. S., Makboul, H. E. & Ali, A. S. (2020). Antimicrobial and antioxidant characteristics of exopolysaccharides produced by Agrobacterium tumefaciens Cs5 and T1. Egyptian Journal of Chemistry, 63(11), 4423-4438. https://doi.org/10.21608/EJCHEM.2020.266 35.2542
  17. Odiba, A.S., Durojaye, O.A., Ezeonu, I.M., Mgbeahuruike, A.C. & Nwanguma, B.C. (2022). A New Variant of Mutational and Polymorphic Signatures in the ERG11 Gene of Fluconazole-Resistant Candida albicans. Infection and Drug Resistance, 15:3111-3133.
  18. Oksel, C., Liyanapathiranage, P., Parajuli, M., Avin, F.A., Jennings, C., Simmons, T. & Baysal-Gurel, F. (2024). Evaluation of Chemical and Biological Products for Control of Crown Gall on Rose. Pathogens, 13(8), 708. https://doi.org/10.3390/pathogens13080708.
  19. Owuama, C.I. (2019). Evaluation of brewing potentials of grains, malts and worts of some sweet sorghum and sorghum varieties. African Journal of Microbiology Research 13(18), 316-322
  20. Peteiro, C. (2018). Alginate Production from Marine Macroalgae, with Emphasis on Kelp Farming. In Alginates and Their Biomedical Applications; Springer: Berlin/Heidelberg, Germany, pp. 27–66.
  21. Prescott, L. M., Harley, J. P. & Klein, D. A. (2011). Microbiology. 8th ed. McGraw Hill, London.
  22. Sandeep, U., Narendra, K., Singh, V. K. & Anshuman, S. (2015).  Isolation, characterization and morphological study of Azotobacter isolates. Journal of Applied and Natural Science, 7 (2), 984– 990.
  23. Tripathi, N. & Sapra, A. (2023). Gram Staining. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan–. PMID: 32965827.
  24. Vatankhah, A., Reezi, S., Izadi, Z., GhasemiVarnamkhasti, M. & Motamedi, A. (2022). Development of an ultrasensitive electrochemical biosensor for detection of Agrobacterium tumefaciens in Rosa hybrida L. Measurement, 187, 110320. https://doi.org/10.1016/j.measurement.2021.110320
  25. Verma, D. K., Niamah, A. K., Patel, A. R., Thakur, M., Sandhu, K. S., Chávez-González, M. L. & Aguilar, C. N. (2020). Chemistry and microbial sources of curdlan with potential application and safety regulations as prebiotic in food and health. Food Research International, 133, 109136. https://doi.org/10.1016/j.foodres.2020.109136
  26. Wang, J., Liu, S., Huang, J., Ren, K., Zhu, Y. & Yang, S. (2023). Alginate: Microbial production, functionalization, and biomedical applications. International Journal of Biological Macromolecules, 242: 125048. https://doi.org/10.1016/j.ijbiomac.2023.125048
  27. Wang, X.Y.Z., Dong, J.J., Xu, G.C., Han, R.Z. & Ni, Y. (2016). Enhanced curdlan production with nitrogen feeding during polysaccharide synthesis by Rhizobium radiobacter. Carbohydr. Polym., 150:385–391
  28. Yang, M., Zhu, Y., Li, Y., Bao, J., Fan, X., Qu, Y., Wang, Y., Hu, Z. & Li, Q. (2016). Production and optimization of curdlan produced by Pseudomonas sp. QL212. Int. J. Biol. Macromol., 89, 25–34.

Curdlan is an important homopolysaccharide, while Alginate is a naturally occurring polymer. Numerous bacteria species have the potential to produce these biopolymers.In this study, Agrobacterium and Azotobacter species capable of producing these polymers were isolated and identified. Soil samples from different legume and cereal farms were inoculated unto mannitol glutamate agar containing aniline blue dyes, Nitrogen free Jensen’s agar medium and Asby agar medium for isolation of the bacteria,The isolates obtained were subjected to biochemical tests and 16S rRNA sequencing for identification. Result showed that 5 species of Agrobaterium grew on aneline blue mannitol glutamate agar (aMGA) exhibiting blue coloration; 2 Azotobacter spp grew on Jensen nitrogen free agar; Agrobaterium and Azotobacter species appeared as Gram negative rods; Agrobaterium spp were KOH sensitive, urease, catalase,citrate positive, hydrolyze starch while Azotobacter spp were positive for methly red, indole, catalase, urease, citrate utilization, starch hydrolysis, and hydrogen sulfide production. Molecular identification showed a band of 1500bp product of 16S rRNA gene of Agrobacterium spp on gel electrophoresis. Result further showed that the Agrobacterium species isolates were capable of producing Curdlan while Azotobacter vinelandii demonstrated Alginate biopolymer production. these findings provide suitable indigenous bacterial strains that will enhance biopolymer production.

Keywords : Agrobacterium, Alginate, Azotobacter, Biopolymers, Curdlan.

Paper Submission Last Date
31 - August - 2026

SUBMIT YOUR PAPER CALL FOR PAPERS
Video Explanation for Published paper

Never miss an update from Papermashup

Get notified about the latest tutorials and downloads.

Subscribe by Email

Get alerts directly into your inbox after each post and stay updated.
Subscribe
OR

Subscribe by RSS

Add our RSS to your feedreader to get regular updates from us.
Subscribe