Levan Produced by the Halophilic Bacterium Bacillus licheniformis BK1 as a Nanoparticle for Protein Immobilization


Ira Oktavia(1), Aidah Nur Fithriah(2), Nur Umriani Permatasari(3), Enny Ratnaningsih(4), Rukman Hertadi(5*)

(1) Biochemistry Research Division, Bandung Institute of Technology, Jl. Ganesa No. 10, Bandung 40132, Indonesia
(2) Biochemistry Research Division, Bandung Institute of Technology, Jl. Ganesa No. 10, Bandung 40132, Indonesia
(3) Biochemistry Research Division, Bandung Institute of Technology, Jl. Ganesa No. 10, Bandung 40132, Indonesia
(4) Biochemistry Research Division, Bandung Institute of Technology, Jl. Ganesa No. 10, Bandung 40132, Indonesia
(5) Biochemistry Research Division, Bandung Institute of Technology, Jl. Ganesa No. 10, Bandung 40132, Indonesia
(*) Corresponding Author


This study examined the potential of levan from the halophilic bacterium Bacillus licheniformis BK1 as a nanoparticle system for protein immobilization. Levan produced by B. licheniformis BK1 was obtained by incubating the bacterium in the optimized Belghith medium, containing 15% (w/v) sucrose, 7.5% (w/v) NaCl and pH 8, in a rotary shaker at 150 rpm for 16 h, at 40 °C. The structure of the levan produced was verified by FTIR and NMR. It appeared that the levan had the same structure as that from Erwinia herbicola. The obtained levan was then used as a nanoparticle system to immobilize BSA and lysozyme proteins. The BSA-nanoparticle had a non-spherical shape with a surface charge of about -4.72 mV and a size distribution in the range of 83–298 nm. In contrast, the lysozyme-nanoparticle exhibited more spherical shapes with a surface charge of -2.57 mV and 206–285 nm size distribution. The efficiency of immobilization was about 74.26% and 81.77% for BSA and lysozyme, respectively. The study thus shows that levan produced by B. licheniformis BK1 can be used as a nanoparticle system for protein immobilization.


levan; levansucrase; lysozyme-nanoparticle; bovine serum albumin-nanoparticle; Bacillus licheniformis BK1

Full Text:

Full Text PDF


[1] Dahech, I., Belghith, K.S., Hamden, K., Feki, A., Belghith, H., and Mejdoub, H., 2011, Antidiabetic activity of levan polysaccharide in alloxan-induced diabetic rats, Int. J. Biol. Macromol., 49 (4), 742–746.

[2] Abdel-Fattah, A.M., Gamal-Eldeen, A.M., Helmy, W.A., and Esawy, M.A., 2012, Antitumor and antioxidant activities of levan and its derivative from the isolate Bacillus subtilis NRC1aza, Carbohydr. Polym., 89 (2), 314–322.

[3] Srikanth, R., Reddy, C.H.S.S.S., Siddartha, G., Ramaiah, M.J., and Uppuluri, K.B., 2015, Review on production, characterization, and applications of microbial levan, Carbohydr. Polym., 120, 102–114.

[4] Srikanth, R., Siddartha, G., Sundhar Reddy, C.H.S.S.S., Harish, B.S., Ramaiah, M.J., and Uppuluri, K.B., 2015, Antioxidant and anti-inflammatory levan produced from Acetobacter xylinum NCIM2526 and its statistical optimization, Carbohydr. Polym., 123, 8–16.

[5] Xu, X., Gao, C., Liu, Z., Wu, J., Han, J., Yan, M., and Wu, Z., 2016, Characterization of the levan produced by Paenibacillus bovis sp. nov BD3526 and its immunological activity, Carbohydr. Polym., 144, 178–186.

[6] Öner, E.T., Hernández, L., and Combie, J., 2016, Review of Levan polysaccharide: From a century of past experiences to future prospects, Biotechnol. Adv., 34 (5), 827–844.

[7] Nakapong, S., Pichyangkura, R., Ito, K., Iizuka, M., and Pongsawasdi, P., 2013, High expression level of levansucrase from Bacillus licheniformis RN-01 and synthesis of levan nanoparticles, Int. J. Biol. Macromol., 54, 30–36.

[8] Ahmed, K.B.A., Kalla, D., Uppuluri, K.B., and Anbazhagan, V., 2014, Green synthesis of silver and gold nanoparticles employing levan, a biopolymer from Acetobacter xylinum NCIM 2526, as a reducing agent and capping agent, Carbohydr. Polym., 112, 539–545.

[9] Bondarenko, O.M., Ivask, A., Kahru, A., Vija, H., Titma, T., Visnapuu, M., Joost, U., Pudova, K., Adamberg, S., Visnapuu, T., and Alamäe, T., 2015, Bacterial polysaccharide levan as stabilizing, non-toxic and functional coating material for microelement-nanoparticles, Carbohydr. Polym., 136, 710–720.

[10] Taran, M., Etemadi, S., and Safaei, M., 2017, Microbial levan biopolymer production and its use for the synthesis of an antibacterial iron(II,III) oxide–levan nanocomposite, J. Appl. Polym. Sci., 134 (12), 44613.

[11] Tabernero, A., González-Garcinuño, Á., Sánchez-Álvarez, J.M., Galán, M.A., and del Valle, E.M.M., 2017, Development of a nanoparticle system based on a fructose polymer: Stability and drug release studies, Carbohydr. Polym., 160, 26–33.

[12] Belghith, K.S., Dahech, I., Belghith, H., and Mejdoub, H., 2012, Microbial production of levansucrase for the synthesis of fructooligosaccharides and levan, Int. J. Biol. Macromol., 50 (2), 451–458.

[13] McBirney, S.E., Trinh, K., Wong-Beringer, A., and Armani, A.M., 2016, Wavelength-normalized spectroscopic analysis of Staphylococcus aureus and Pseudomonas aeruginosa growth rates, Biomed. Opt. Express, 7 (10), 4034–4042.

[14] Sezer, A.D., Kazak, H., Öner, E.T., and Akbua, J., 2011, Levan-based nanocarrier system for peptide and protein drug delivery: Optimization and influence of experimental parameters on the nanoparticle characteristics, Carbohydr. Polym., 84 (1), 358–363.

[15] Sezer, A.D., Sarılmışer, H.K., Rayaman, E., Çevikbaş, A., Öner, E.T., and Akbuğa, J., 2017, Development and characterization of vancomycin-loaded levan-based microparticular system for drug delivery, Pharm. Dev. Technol., 22 (5), 627–634.

[16] Mamay, Wahyuningrum, D., and Hertadi, R., 2015, Isolation and characterization of levan from moderate halophilic bacteria Bacillus licheniformis BK AG21, Procedia Chem., 16, 292–298.

[17] Zhang, T., Li, R., Qian, H., Mu, W., Miao, M., and Jiang, B., 2014, Biosynthesis of levan by levansucrase from Bacillus methylotrophicus SK 21.002, Carbohydr. Polym., 101, 975–981.

[18] Li, W., Yu, S., Zhang, T., Jiang, B., and Mu, W., 2015, Recent novel applications of levansucrases, Appl. Microbiol. Biotechnol., 99 (17), 6959–6969.

[19] Ebel, C., Madern, D., and Zaccai, G., 2009, “Molecular Adaptation of Halophilic Proteins” in Extremophiles, vol. II, Eds., Gerday, C., and Glandsdorff, N., Eolss Publisher Co. Ltd., Oxford, England.

DOI: https://doi.org/10.22146/ijc.41064

Article Metrics

Abstract views : 2093 | views : 2151

Copyright (c) 2020 Indonesian Journal of Chemistry

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.


Indonesian Journal of Chemistry (ISSN 1411-9420 / 2460-1578) - Chemistry Department, Universitas Gadjah Mada, Indonesia.

Analytics View The Statistics of Indones. J. Chem.