Isolation of Biofilm-Bioflocculant Producing Bacteria from Palm Oil Mill Effluent


  • Wan Rosmiza Zana Wan Dagang Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia
  • Nuraihanah Najihah Zaidi Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia



Biofilm, bioflocculant, palm oil mill effluent, bacteria


A variety of flocculants have been used to treat wastewater by agglomerating various colloidal substances and cells. Due to the adverse effects and high cost of conventional flocculants, bioflocculant which are originally produced by microorganisms are gaining attention. Therefore, this study aimed to identify the biofilm-bioflocculant-producing bacteria which was isolated from palm oil mill effluent in Johor, Malaysia. The isolated bacteria were identified by their colony morphology, Gram-staining and 16S rRNA gene sequence analysis. All isolates were subjected to bioflocculant assay with kaolin clay suspension as a model of wastewater to assess the flocculation efficiency of the bacterial bioflocculants. Two different isolates were successfully identified as Gram-negative bacteria Stenotrophomonas koreensis TR6-01 and Alishewanella agri BL06. S. koreensis TR6-01 shows the highest flocculating activity at 82.79%, followed by A. agri BL06 with 78.52%, respectively. The strongest biofilm producer was shown by S. koreensis TR6-01 and thus, indicates good candidate for future wastewater treatment technology.

Author Biography

Nuraihanah Najihah Zaidi, Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia




Peng, L., Yang, C., Zeng, G., Wang, L., Dai, C., Long, Z., Liu, H. and Zhong, Y. (2014). Characterization and application of bioflocculant prepared by Rhodococcus erythropolis using sludge and livestock wastewater as cheap culture media. Applied Microbiology and Biotechnology, 98, 6847-6858.

Makapela, B., Okaiyeto, K., Ntozonke, N., Nwodo, U. U., Green, E., Mabinya, L. V., & Okoh, A. I. (2016). Assessment of Bacillus pumilus isolated from fresh water milieu for bioflocculant production. Applied Sciences, 6(8), 211.

Tsilo, P. H., Basson, A. K., Ntombela, Z. G., Maliehe, T. S., & Pullabhotla, V. R. (2022). Production and characterization of a bioflocculant from Pichia kudriavzevii MH545928. 1 and Its Application in Wastewater Treatment. International Journal of Environmental Research and Public Health, 19(6), 3148.

Abbas, S.Z., Yong, Y.C., Ali Khan, M., Siddiqui, M.R., Hakami, A.A.H., Alshareef, S.A., Otero, M. and Rafatullah, M. (2020). Bioflocculants produced by bacterial strains isolated from palm oil mill effluent for application in the removal of Eriochrome Black T dye from water. Polymers, 12(7), 1545.

Al-Amshawee, S. K., Yunus, M. Y., & Azoddein, A. A. (2020). A review study of biofilm bacteria and microalgae bioremediation for palm oil mill effluent: possible approach. IOP Conference Series: Materials Science and Engineering, 736 (2), 022034.

Aryal S. Gram Staining: Principle, Procedure, Interpretation, Examples and Animation 2022. (accessed March 20, 2023).

Ndejiko, J. M., & Dagang, W. R. Z. W. (2019). Flocculation behaviour of bioflocculant produced from chicken viscera. E3S Web of Conferences, 90, 01013.

Saggu, S. K., Jha, G., & Mishra, P. C. (2019). Enzymatic degradation of biofilm by metalloprotease from Microbacterium sp. SKS10. Frontiers in Bioengineering and Biotechnology, 7, 192.

Molobela, I. P., Cloete, T. E., & Beukes, M. (2010). Protease and amylase enzymes for biofilm removal and degradation of extracellular polymeric substances (EPS) produced by Pseudomonas fluorescens bacteria. 10 (4), 1515–24.

Stepanović, S., Vuković, D., Hola, V., Bonaventura, G. D., Djukić, S., Ćirković, I., & Ruzicka, F. (2007). Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. Apmis, 115(8), 891-899.

He, C. Q., Meng, S. L., Yan, H. Y., Ding, N. Z., He, H. B., Yan, J. X., & Xu, G. L. (2012). Isolation and identification of a novel rabies virus lineage in China with natural recombinant nucleoprotein gene. PLoS One, 7(12), e49992.

Tamura, K., Stecher, G., Peterson, D., Filipski, A., & Kumar, S. (2013). MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular biology and evolution, 30(12), 2725-2729.

Ahern H. Microbiology: a laboratory experience. New York: Open SUNNY Textbooks, Milne Library; 2018.

Fazil, N. A., Alam, M. Z., Azmi, A. S., & Mansor, M. F. (2018). Isolation and screening of bacteria with biofilm formation ability and characterization with hydrolytic enzyme production for enhanced biogas production 96-101. Malaysian Journal of Microbiology, 96-101.

Yang, H. C., Im, W. T., Kang, M. S., Shin, D. Y., & Lee, S. T. (2006). Stenotrophomonas koreensis sp. nov., isolated from compost in South Korea. International journal of systematic and evolutionary microbiology, 56(1), 81-84.

Kim, M. S., Jo, S. K., Roh, S. W., & Bae, J. W. (2010). Alishewanella agri sp. nov., isolated from landfill soil. International journal of systematic and evolutionary microbiology, 60(9), 2199-2203.

Félicité, D.N., Daïna, N.T., Roméo, F.T., Joël, T.S., Fatima, N., Joël, N.N., Yanick, K., Josiane, B., Felix, F., Mpondo, E.M. and Beng, V.P. (2021). Isolation of Bacteria with Purifying Potential and Application in the Treatment of Effluents from an Artisanal Palm Oil Mill in the Littoral Region of Cameroon. Journal of Environmental Protection, 12:462–71.

Chen, H., Zhong, C., Berkhouse, H., Zhang, Y., Lv, Y., Lu, W., Yang, Y. and Zhou, J. (2016). Removal of cadmium by bioflocculant produced by Stenotrophomonas maltophilia using phenol-containing wastewater. Chemosphere, 155, 163-169.

Azimi, A., Aslanimehr, M., Yaseri, M., Shadkam, M., & Douraghi, M. (2020). Distribution of smf-1, rmlA, spgM and rpfF genes among Stenotrophomonas maltophilia isolates in relation to biofilm-forming capacity. Journal of Global Antimicrobial Resistance, 23, 321-326.

Passerini de Rossi, B., Calenda, M., Vay, C., & Franco, M. (2007). Biofilm formation by Stenotrophomonas maltophilia isolates from device-associated nosocomial infections. Revista Argentina de microbiología, 39(4), 204-212.

Mohammed, J. N., & Wan Dagang, W. R. Z. (2019). Culture optimization for production and characterization of bioflocculant by Aspergillus flavus grown on chicken viscera hydrolysate. World Journal of Microbiology and Biotechnology, 35, 1-19.




How to Cite

Wan Dagang, W. R. Z., & Zaidi, N. N. (2023). Isolation of Biofilm-Bioflocculant Producing Bacteria from Palm Oil Mill Effluent. Journal of Materials in Life Sciences (JOMALISC), 2(1), 115–122.