Production of Polyhydroxyalkanoates (PHAs) from Oil Palm Waste Using Paraburkholderia sp. PFN29
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Abstract
The increasing accumulation of plastic waste has intensified efforts to identify sustainable biodegradable alternatives. Polyhydroxyalkanoates (PHAs) are microbial polyesters with properties similar to conventional plastics; however, their commercial feasibility is limited by high production costs. This study investigated the use of oil palm waste, specifically palm kernel meal (PKM) and empty fruit bunch (EFB), as feedstocks for PHA production by Paraburkholderia sp. PFN29. Biomass pretreatment used sodium hydroxide (1-5% w/v) followed by hydrogen peroxide (3% v/v), and enzymatic hydrolysis with pectinase, xylanase, and cellulase. Reducing sugars were quantified using the dinitrosalicylic acid (DNS) method, and morphological changes were examined via scanning electron microscopy (SEM). Optimal saccharification with 3% (w/v) NaOH at 50°C for 48 h yielded 10.53±0.33 and 21.02±0.38 mg/mL reducing sugars from PKM and EFB, respectively. Glucose was the predominant sugar in both hydrolysates. PHA biosynthesis was assessed under various carbon- and nitrogen-supplementation conditions. The EFB hydrolysate at 100 ml supplemented with 0.1 g NH₄Cl produced the highest PHA concentration (1.12±0.02 g/L), PHA content (59.02%), and productivity (0.01 g/L/h), which was comparable to that of the mineral medium control. The PKM hydrolysate supported optimal PHA production without supplementation (PHA content, 39.02%). Fourier-transform infrared (FTIR) and nuclear magnetic resonance (NMR) analyses confirmed PHB-type PHA production. These findings indicate that palm oil residues, particularly EFB hydrolysates under optimized nitrogen conditions, are promising substrates for sustainable PHA production.
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References
Chae, Y., & An, Y.-J. (2018). Current research trends on plastic pollution and ecological impacts on the soil ecosystem: A review. Environmental Pollution, 240, 387-395. https://doi.org/10.1016/j.envpol.2018.05.008
Chakrawal, A., Calabrese, S., Herrmann, A. M., & Manzoni, S. (2022). Interacting bioenergetic and stoichiometric controls on microbial growth. Frontiers in Microbiology, 13, Article 859063. https://doi.org/10.3389/fmicb.2022.859063
Chen, R., Wang, Y.-Z., Liao, Q., Zhu, X., & Xu, T.-F. (2013). Hydrolysates of lignocellulosic materials for biohydrogen production. BMB Reports, 46(5), 244-251. https://doi.org/10.5483/BMBRep.2013.46.5.038
Cui, T.-W., Shi, Y.-P., & Gong, X.-Y. (2017). Effects of C/N in the substrate on the simultaneous production of polyhydroxyalkanoates and extracellular polymeric substances by Haloferax mediterranei via kinetic model analysis. RSC Advances, 7, 18953-18961. https://doi.org/10.1039/C7RA02131C
da Silva, M. B., de Lima Araújo, R. R., Almeida, R. M. R. G., de Farias Silva, C. E., Brandão, M.R.P., de Menezes Bernardino, T., Lôbo, L. N., de Freitas, J. M. D., & de Freitas, J. D. (2025). Evaluating sodium hydroxide and hydrogen peroxide as chemical treatment for cellulose extraction from Clitoria fairchildiana pruning residues. Reactions, 6(4), Article 60. https://doi.org/10.3390/reactions6040060
Dimawarnita, F., Faramitha, Y., Kalbuadi, D. N., Prakoso, H. T., Puspitasari, I., & Prasetyo, D. (2023). Characterization of cellulose from oil palm empty fruit bunches by fast delignification process with different solvents. Menara Perkebunan, 91(2), 96-105.
Díez, D., Urueña, A., Piñero, R., Barrio, A., & Tamminen, T. (2020). Determination of hemicellulose, cellulose, and lignin content in different types of biomasses by thermogravimetric analysis and pseudocomponent kinetic model (TGA-PKM method). Processes, 8(9), Article 1048. https://doi.org/10.3390/pr8091048
Elgharbawy, A. A., Alam, M. Z., Moniruzzaman, M., Kabbashi, N. A., & Jamal, P. (2018). Chemical and structural changes of pretreated empty fruit bunch (EFB) in ionic liquid-cellulase compatible system for fermentability to bioethanol. 3 Biotech, 8(5), Article 236. https://doi.org/10.1007/s13205-018-1253-8
Etxabide, A., Kilmartin, P. A., Guerrero, P., de la Caba, K., Hooks, D. O., West, M., & Singh, T. (2022). Polyhydroxybutyrate (PHB) produced from red grape pomace: Effect of purification processes on structural, thermal and antioxidant properties. International Journal of Biological Macromolecules, 217, 449-456. https://doi.org/10.1016/j.ijbiomac.2022.07.072
Fayshal, M. A. (2024). Current practices of plastic waste management, environmental impacts, and potential alternatives for reducing pollution and improving management. Heliyon, 10(23), Article e40838. https://doi.org/10.1016/j.heliyon.2024.e40838
Gao, Q., Yang, H., Wang, C., Xie, X.-Y., Liu, K.-X., Lin, Y., Han, S.-Y., Zhu, M., Neureiter, M., Lin, Y., & Ye, J.-W. (2022). Advances and trends in microbial production of polyhydroxyalkanoates and their building blocks. Frontiers in Bioengineering and Biotechnology, 10, Article 966598. https://doi.org/10.3389/fbioe.2022.966598
Ghazali, N. F., & Makhtar, N. A. (2018). Enzymatic hydrolysis of oil palm empty fruit bunch and its kinetics. Malaysian Journal of Analytical Sciences, 22(4), 715-722. https://doi.org/10.17576/mjas-2018-2204-18
Hahn, S. K., Chang, Y. K., Kim, B. S., & Chang, H. N. (1994). Optimization of microbial poly(3-hydroxybutyrate) recovery using dispersions of sodium hypochlorite solution and chloroform. Biotechnology and Bioengineering, 44(2), 256-261. https://doi.org/10.1002/bit.260440215
IUCN. (2024). Plastic pollution and its impacts on ecosystems and human health. International Union for Conservation of Nature.
Irfan, M., Asghar, U., Nadeem, M., Nelofer, R., & Syed, Q. (2016). Optimization of process parameters for xylanase production by Bacillus sp. in submerged fermentation. Journal of Radiation Research and Applied Sciences, 9(2), 139-147. https://doi.org/10.1016/j.jrras.2015.10.008
Iyayi, E. A., Agboola, A. F., & Baah, J. (2015). Nutritional value of palm kernel cake and meal for livestock and fish feeding: A review. Tropical Animal Production Investigation, 18(2), 54-66.
Jung, J. Y., Ha, S. Y., & Yang, J.-K. (2022). Comparison of carbohydrate composition in lignocellulosic biomass by high performance liquid chromatography and gas chromatography analysis. BioResources, 17(1), 1454-1466. https://doi.org/10.15376/biores.17.1.1454-1466
Justes, E., Mary, B., & Nicolardot, B. (2009). Quantifying and modelling C and N mineralization kinetics of catch crop residues in soil: Parameterization of the residue decomposition module of STICS model for mature and non-mature residues. Plant and Soil, 325(1-2), 171-189. https://doi.org/10.1007/s11104-009-9966-4
Kim, S. (2018). Enhancing bioethanol productivity using alkali-pretreated empty palm fruit bunch fiber hydrolysate. BioMed Research International, 2018, Article 5272935. https://doi.org/10.1155/2018/5272935
Li, Z., Yang, J., & Loh, X. J. (2016). Polyhydroxyalkanoates: Opening doors for a sustainable future. NPG Asia Materials, 8, Article e265. https://doi.org/10.1038/am.2016.48
Lu, H., Sato, H., & Kazarian, S. G. (2021). Visualization of inter- and intramolecular interactions in poly(3-hydroxybutyrate)/poly(L-lactic acid) (PHB/PLLA) blends during isothermal melt crystallization using attenuated total reflection fourier transform infrared (ATR FT-IR) spectroscopic imaging. Applied Spectroscopy, 75(8), 980-987. https://doi.org/10.1177/00037028211010216
Lyu, Q., Dar, R. A., Baganz, F., Smoliński, A., Rasmey, A.-H. M., Liu, R., & Zhang, L. (2025). Effects of lignocellulosic biomass-derived hydrolysate inhibitors on cell growth and lipid production during microbial fermentation of oleaginous microorganisms—A review. Fermentation, 11(3), Article 121. https://doi.org/10.3390/fermentation11030121
Miller, G. L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31(3), 426-428. https://doi.org/10.1021/ac60147a030
Østby, H., Hansen, L. D., Horn, S. J., Eijsink, V. G. H., & Várnai, A. (2020). Enzymatic processing of lignocellulosic biomass: Principles, recent advances and perspectives. Journal of Industrial Microbiology and Biotechnology, 47(9), 623-657. https://doi.org/10.1007/s10295-020-02301-8
Palamae, S., Dechatiwongse, P., Choorit, W., Chisti, Y., & Prasertsan, P. (2017). Cellulose and hemicellulose recovery from oil palm empty fruit bunch (EFB) fibers and production of sugars from the fibers. Carbohydrate Polymers, 155, 491-497. https://doi.org/10.1016/j.carbpol.2016.09.004
Pant, S., Ritika, Komesu, A., Penteado, E. D., Diniz, A. A. R., Rahman, M. A., & Kuila, A. (2021). NaOH pretreatment and enzymatic hydrolysis of Brassica juncea using mixture of cellulases. Environmental Technology and Innovation, 21, Article 101324. https://doi.org/10.1016/j.eti.2020.101324
Premjet, D., & Premjet, S. (2025). Enhanced sugar and bioethanol production from broom grass via NaOH-autoclave pretreatment. Polymers, 17(3), Article 266. https://doi.org/10.3390/polym17030266
R Core Team. (2016). R: A language and environment for statistical computing. R Foundation for Statistical Computing. https://www.R-project.org/
Radzi, N. A. M., Sofian, A. H., & Jamari, S. S. (2020). Structural studies of surface modified oil palm empty fruit bunch with alkaline pre-treatment as a potential filler for the green composite. Jurnal Tribologi, 26, 75-83.
Said, F. M., Hamid, N. F., Razali, M. A.-A., & Daud, N. F. S. (2021). Lignocellulosic of oil palm biomass to chemical product via fermentation. In H. Kamyab (Ed.). Elaeis guineensis. IntechOpen. https://doi.org/10.5772/intechopen.92931
Samrot, A. V., Samanvitha, S. K., Shobana, N., Renitta, E. R., Senthilkumar, P., Kumar, S. S., Abirami, S., Dhiva, S., Bavanilatha, M., Prakash, P., Saigeetha, S., Shree, K. S., & Thirumurugan, R. (2021). The synthesis, characterization and applications of polyhydroxyalkanoates (PHAs) and PHA-based nanoparticles. Polymers, 13(19), Article 3302. https://doi.org/10.3390/polym13193302
Sartori, T., Tibolla, H., Prigol, E., Colla, L. M., Costa, J. A. V., & Bertolin, T. E. (2015). Enzymatic saccharification of lignocellulosic residues by cellulases obtained from solid state fermentation using Trichoderma viride. BioMed Research International, 2015, Article 342716. https://doi.org/10.1155/2015/342716
Satapathy, S., Rout, J. R., Kerry, R. G., Thatoi, H., & Sahoo, S. L. (2020). Biochemical prospects of various microbial pectinase and pectin: An approachable concept in pharmaceutical bioprocessing. Frontiers in Nutrition, 7, Article 117. https://doi.org/10.3389/fnut.2020.00117
Shen, Y., Peng, H., & Bi, H. (2025). Insight into the physicochemical characteristics and biological features of dietary polysaccharides extracted from palm kernel cake. Grain and Oil Science and Technology, 8(2), 77-88. https://doi.org/10.1016/j.gaost.2025.03.002
Sehgal, R., & Gupta, R. (2020). Polyhydroxyalkanoate and its efficient production: An eco-friendly approach towards development. 3 Biotech, 10(12), Article 549. https://doi.org/10.1007/s13205-020-02550-5
Shin, N., Kim, S. H., Oh, J., Kim, S., Lee, Y., Shin, Y., Choi, S., Bhatia, S. K., Jeon, J.-M., Yoon, J.-J., Joo, J. C., & Yang, Y.-H. (2024). Evaluation of blended poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) properties containing various 3HHx monomers. Polymers, 16(21), Article 3077. https://doi.org/10.3390/polym16213077
Sriyapai, T., Chuarung, T., Kimbara, K., Samosorn, S., & Sriyapai, P. (2022). Production and optimization of polyhydroxyalkanoates (PHAs) from Paraburkholderia sp. PFN 29 under submerged fermentation. Electronic Journal of Biotechnology, 56, 1-11. https://doi.org/10.1016/j.ejbt.2021.12.003
Sruthi, N. U., Rao, P. S., Bennett, S. J., & Bhattarai, R. R. (2023). Formulation of a synergistic enzyme cocktail for controlled degradation of sorghum grain pericarp. Foods, 12(2), Article 306. https://doi.org/10.3390/foods12020306
Sun, S., Yang, S., Qiu, Y., Ding, J., Wang, W., Wu, F., & Chen, G. Q. (2025). Life cycle design of polyhydroxyalkanoates (PHA). National Science Review, 12(12), Article nwaf517. https://doi.org/10.1093/nsr/nwaf517
Sundalian, M., Larissa, D., & Suprijana, O. (2021). Contents and utilization of palm oil fruit waste. Biointerface Research in Applied Chemistry, 11(3), 10148-10160. https://doi.org/10.33263/BRIAC113.1014810160
Tang, X., Liu, X., Zhang, Y., Peng, P. & Hang, H. (2021). The nutritive value of palm kernel cake and its application in low quality diets of broiler chickens. Pakistan Journal of Agricultural Sciences, 58(5), 1429-1436. https://doi.org/10.21162/pakjas/21.1187
Tareen, A. K., Punsuvon, V., & Parakulsuksatid, P. (2020). Investigation of alkaline hydrogen peroxide pretreatment to enhance enzymatic hydrolysis and phenolic compounds of oil palm trunk. 3 Biotech, 10(4), Article 179. https://doi.org/10.1007/s13205-020-02169-6
Thanapimmetha, A., Khomlaem, C., Saisriyoot, M., Naktham, N., & Srinophakun, P. (2023). Improved bioethanol production from oil palm empty fruit bunch using different fermentation strategies. Agriculture and Natural Resources, 57(4), 689-696. https://doi.org/10.34044/j.anres.2023.57.4.13
Trakunjae, C., Boondaeng, A., Apiwatanapiwat, W., Kosugi, A., Arai, T., Sudesh, K., & Vaithanomsat, P. (2021). Enhanced polyhydroxybutyrate (PHB) production by newly isolated rare actinomycetes Rhodococcus sp. strain BSRT1-1 using response surface methodology. Scientific Reports, 11, Article 1896. https://doi.org/10.1038/s41598-021-81386-2
Ulia, H., Samah, S. D., & Nurmalasari, E. (2025). The production of polyhydroxyalkanoate (PHA) bioplastic from palm oil mill effluent (POME) using Pseudomonas aeruginosa. Jurnal Kimia Sains dan Aplikasi, 28(7), 387-395. https://doi.org/10.14710/jksa.28.7.387-395
Wahyudin, C. I., & Oge, L. (2025). Utilization of oil palm waste as a renewable energy source: A current literature review. Journal of Agriculture, Agribusiness, Welfare, Technology, Humanity, Environment, Social, and Economy, 1(2), 70-79.
Wang, W., Wang, X., Zhang, Y., Yu, Q., Tan, X., Zhuang, X., & Yuan, Z. (2020). Effect of sodium hydroxide pretreatment on physicochemical changes and enzymatic hydrolysis of herbaceous and woody lignocelluloses. Industrial Crops and Products, 145, Article 112145. https://doi.org/10.1016/j.indcrop.2020.112145
Xia, J., Shu, J., Yao, K., Xu, J., Yu, X., Xue, X., Ma, D., & Lin, X. (2020). Synergism of cellulase, pectinase and xylanase on hydrolyzing differently pretreated sweet potato residues. Preparative Biochemistry and Biotechnology, 50(2), 181-190. https://doi.org/10.1080/10826068.2019.1680390
Yustinah, Hidayat, N., Alamsyah, R., Roslan, A. M., Hermansyah, H., & Gozan, M. (2019). Production of polyhydroxybutyrate from oil palm empty fruit bunch (OPEFB) hydrolysates by Bacillus cereus suaeda B-001. Biocatalysis and Agricultural Biotechnology, 18, Article 101019. https://doi.org/10.1016/j.bcab.2019.01.057
Zhang, L., Jiang, Z., Tsui, T-H, Loh, K-C, Dai, Y., & Tong, Y. W. (2022). A review on enhancing Cupriavidus necator fermentation for poly(3-hydroxybutyrate) (PHB) production from low-cost carbon sources. Frontiers in Bioengineering and Biotechnology, 10, Article 946085. https://doi.org/10.3389/fbioe.2022.946085
Zhang, Y., Sun, W., Wang, H., & Geng, A. (2013). Polyhydroxybutyrate production from oil palm empty fruit bunch using Bacillus megaterium R11. Bioresource Technology, 147, 307-314. https://doi.org/10.1016/j.biortech.2013.08.029
Zubaidah, S., Hanim, C., Ariyadi, B., Baskara, A. P., & Zuprizal. (2024). Nutrient composition and cell-wall structure of palm kernel cake supplemented with enzymes. Advances in Animal and Veterinary Sciences, 12(6), 1191-1198. https://dx.doi.org/10.17582/journal.aavs/2024/12.6.1191.1198
Zytner, P., Kumar, D., Elsayed, A., Mohanty, A., Ramarao, B. V., & Misra, M. (2023). A review on polyhydroxyalkanoate (PHA) production through the use of lignocellulosic biomass. RSC Sustainability, 1, 2120-2134. https://doi.org/10.1039/D3SU00126A
