Harnessing molasses as a low-cost carbon source for production of poly-hydroxy butyrate (PHB) using Burkholderia sp. B73 bacteria


  • Diah - RATNANINGRUM Research for Clean Technology - Indonesian Institute of Science http://orcid.org/0000-0001-6946-1372
  • Een Sri ENDAH Research for Clean Technology - Indonesian Institute of Science
  • Puspita LISDIYANTI Research Centre for Biotechnology-Indonesian Institute of Science
  • Sri PRIATNI Research for Clean Technology - Indonesian Institute of Science
  • Vienna SARASWATY Research for Clean Technology - Indonesian Institute of Science




C/N, ratio, sugarcane


Burkholderia sp. has been reported as a poly-hydroxy-butyrate (PHB) producer. PHB is a natural polyester class with a wide range of applications in foods, medicines, and biomedicines. However, the high production cost of PHB may limit its potential. Molasses, a by-product of the sugarcane industry available abundantly, may be used as an alternative carbon source of PHB production. In this research, we aimed to evaluate PHB production by Burkholderia sp. B73 in fermentation media using molasses as an alternative carbon source. Small-scale experiments were performed in Erlenmeyer flasks on a shaker at 150 rpm and 30 °C to evaluate the best initial C/N ratio for biomass accumulation and PHB production. A set of parameters including bacterial growth, dry cell weight, yield, and FTIR spectrum of PHB were observed.  The results showed that molasses could be used to grow Burkholderia sp. B73 and the highest PHB production was obtained when a 20:1 C/N ratio of molasses was applied in the fermentation medium. In addition, when the initial pH was adjusted to 7.0, the highest PHB yield was also produced. More importantly, the use of molasses as a carbon source improved the PHB yield by nearly 2-fold compared with our previous report using a synthetic Ramsay’s minimal medium. In conclusion, the experiment results showed that molasses could be used as a low-cost carbon source for PHB production by Burkholderia sp. B73 bacteria.


Download data is not yet available.

Author Biographies

Diah - RATNANINGRUM, Research for Clean Technology - Indonesian Institute of Science


First Researcher

Department Product and Clean Product

Een Sri ENDAH, Research for Clean Technology - Indonesian Institute of Science


Puspita LISDIYANTI, Research Centre for Biotechnology-Indonesian Institute of Science

Madya Researcher

Sri PRIATNI, Research for Clean Technology - Indonesian Institute of Science

Madya Researcher

Vienna SARASWATY, Research for Clean Technology - Indonesian Institute of Science

First Researcher


Accorsi R, Cascini A, Cholette S, Manzini R & Mora C (2014). Economic and environmental assessment of reusable plastic containers: A food catering supply chain case study. Intl J of Production Economics 152, 88-101.

Aramvash A, Moazzeni ZF & Gholami BN (2018). Comparison of different solvents for extraction of polyhydroxybutyrate from Cupriavidus necator. Eng in Life Sci 18, 20-28.

Ashraf S, Ali S & Ikramul H (2015). Pre-treatment of raw sugarcane molasses by metal complexing agents for improved citric acid fermentation by Aspergillus niger. Intl J of Res in Pharm and Biosci 2(6), 34-40.

Atifah N, Khawar S & A Suryani (2007). Kajian fermentasi bioplastik poli-(3-Hidroksialkanoat)/PHA oleh Ralstonia etropha menggunakan sumber karbon hidrolisat pati sagu. Jurnal teknologi Pertanian 8(3), 160-171

Beaulieu M, Y Beaulieu, J Melinard, S Pandian & J Goulet (1995). Influence of ammonium salts and cane molasses on growth of Alcaligenes eutrophus and production of polyhydroxybutyrate. Appl and Environmental Microbiology 61(1), 165-169.

Bhagowati P, Pradhan S, Dash HR & Das S (2015). Production, optimization and characterization of polyhydroxybutyrate, a biodegradable plastic by Bacillus spp. Biosci Biotechnol Biochem 79, 1454–63

Bhuwal, Anish K, G Singh, NK Aggarwal, V Goyal & A Yadav (2013). Isolation and screening of polyhydroxyalkanoates producing bacteria from pulp, paper, and cardboard industry wastes. International Journal of Biomaterials. https://doi.org/10.1155/2013/752821.

Botha T & H von Blottnitz (2006). A comparison of the environmental benefits of bagasse-derived electricity and fuel ethanol on a life-cycle basis. Energy Policy 34(17), 2654-2661.

Chauhan MK, Varun, S Chaudhary S, S Kumar & Samarm (2011). Life cycle assessment of sugar industry: A review. Renewable and Sustainable Energy Reviewers. https://doi.org/10.1016/j.rser.2011.04.033.

Choi J & Lee S (1999). Factors affecting the economics of polyhydroxyalkanoate production by bacterial fermentation. Appl Microbiol and Biotech 51, 13–21.

Gomaa EZ (2014). Production of polyhydroxyalkanoates (PHAs) by Bacillus subtilis and Escherichia coli grown on cane molasses fortified with ethanol. Braz Arch of Biol and Tech 57(1), 145-154.

Gouda MK, Azza ES & Sanaa HO (2001). Production of PHB by a Bacillus megaterium strain using sugarcane molasses and corn steep liquor as sole carbon and nitrogen sources. Microbiol Res 156, 201-207.

Hassan MA, Elsayed KB, Salah GA & Hussien RH (2016). Production and characterization of polyhydroxybutyrate (PHB) produced by Bacillus sp. isolated from Egypt. Journal of Applied Pharmaceutical Science 6 (4), 46-51. DOI: 10.7324/JAPS.2016.60406

Irwandi, Djamaan A & Agustien A (2018). Pengaruh konsentrasi minyak kelapa sawit mentah terhadap jumlah biomassa bakteri Baciluss spp. penghasil polimer poli (3-hidroksibutirat). Scientia Jurnal Farmasi dan Kesehatan 8(1), 64-72

Jiun YC, Tan Y, Samian MR & Sudesh K (2010). Isolation and characterization of Burkholderia sp. USM (JCM15050) capable of producing polyhydroxyalkanoate (PHA) from triglycerides, fatty acids and glycerols. J Polym Environ 18,584–592. DOI 10.1007/s10924-010-0204-1

Kamravamanesh D, Lackner M & Herwig C (2018). Bioprocess engineering aspects of sustainable polyhydroxyalkanoate production in cyanobacteria. Bioeng (Basel) 5(4), 111.

Khleifat K (2007). Effect of substrate adaptation, carbon starvation and cell density on the biodegradation. of phenol by Actinobacillus sp. Fresenius Environmental Bulletin 16, 726-730.

Koller M (2017). Advances in polyhydroxyalkanoate (PHA) production. Bioengineering (Basel) 4(4), 88.

Kresnawaty I, Mulyatni AS, Eris DD & Prakoso HT (2016). Characterization of PHA produced by Pseudomonas aeruginosa and Bacillus subtilis inoculated in palm oil mill effluent (POME) media. Menara Perkebunan 82(2), 57-63.

Lee SN, Moon YL & WH Park (2002). Thermal stabilization of poly(3-hydroxybutyrate) by Poly (glycidyl methacrylate) J Appl Polym Sci. 83 2945–52

Luo, Rong C, Yun LW, & Mohamed EN (2014). The industrial production of PHA. In Polyhydroxyalkanoates (PHAs): Biosynthesis, Industrial Production and Applications in Medicine. Chapter 13. Nanotechnology Science &Technology Inc. New York 11788-3619, USA

Marangoni C, A Furigo Jr & Aragão GMF (2001). The influence of substrate source on the growth of Ralstonia eutropha, aiming at the production of polyhydroxy alkanoate. Braz J of Chem Eng 18(2), 175-180.

Matias F, CA Brandt, ES de Silva & MF de Adrade Rodrigues (2017). Polyhydroxybutyrate and polyhydroxydodecanoate produced by Burkholderia contaminans IPT553. Journal Appl Microbiol 123 (1),124-133.

Mostafa NA, Farag AA, Abodief HM & Tayeb AM (2018). Production of biodegradable plastic from agricultural wastes. Arab J of Chem 11(4), 546–553.

Naheed N & Jamil N (2014). Optimization of biodegradable plastic production on sugar cane molasses in Enterobacter sp. SEL2. Braz J of Microbiol 45(2 ), 417-426.

Oliveira FC, Denisse MGF & L Castilho (1999). Production of poly(3-hydroxybutyrate) by solid-state fermentation with Ralstonia eutropha. Biotechnology letters 26 (24): 1851-5. DOI:10.1007/s10529-004-5315-0

Pan W, Perrotta JA, Stipanovic AJ, Nomura CT & Nakas JP (2012). Production of polyhydroxyalkanoates by Burkholderia cepacia ATCC 17759 using a detoxified sugar maple hemicellulosic hydrolysate. J of Ind Microbiol and Biotech 39(3), 459-469.

Penkhrue W, Jendrossek D, Khanongnuch C, Pathom AW, Aizawa T, Behrens RL & Lumyong S (2020). Response surface method for polyhydroxybutyrate (PHB) bioplastic accumulation in Bacillus drentensis BP17 using pineapple peel. PLoS One 15(3): e 0230443.

Ratnaningrum D, Een SE, Akbar HDA, Vienna S, Puspita L, Eva Frasnawaty & Sri P (2020). The effect of inoculum and glucose addition of polyhydroxyalkanoate production by Brevibacterium sp. B45. Menara Perkebunan 89 (1), 1-7.

Ratnaningrum D, V Saraswaty, S Priatni, Puspita L, A Purnomo & S Pudjiraharti (2019). Screening of polyhydroxyalkanoates (PHA)-producing bacteria from soil bacteria strains. IOP Conf. Series: Earth and Environmental Science 277.

Saleem, Faiza, Reema A, Yasar S, Shagufta N, Q Syed, Syed Q, N Munir, Nazia K & Abdul RK (2014). Analysis and Evaluation of Growth Parameters for Optimum Production of Polyhydroxybutyrate (PHB) by Bacillus thuringiensis strain CMBL-BT-6. Pak J of Zoo 46(5), 1337-1344

Senthilkumar S, Suganya T, Deepa K, Muralidharan J & Sasikala K (2016). Supplementation of molasses in livestock feed. International Journal of Science, Environment and Technology, 5 (3): 1243 – 1250. ISSN 2278-3687

Spiekerman P Rehm B, Kalscheuer RBD & Steinbüchel A (1999). A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds. Arch. of Microbiol 171, 73–80

Sudesh K, Abe H & Doi Y (2000). Synthesis, structure and properties of Polyhydroxyalkanoates: biological polyesters. Prog in Polymer Sci 25, 1503-1555.

Webb HK, Jaims ARJC & Elena PI (2013). Plastic degradation and its environmental implications with special reference to poly(ethylene terephthalate). Polymers 5, 1-18.

Wen Z, Wei L & Shulin Ch (2004). Hydrolysis of animal manure lignocellulosics for reducing sugar production. Biores Tech 91(1), 31-39.

Wu Q, H Huang, G Hu, J Chen, KP Ho & GQ Chen (2001). Production of poly-3-hydroxybutrate by Bacillus Sp. JMa5 cultivated in molasses media. Antonie van Leeuwenhoek, International Journal of General and Mol Microbiol 80, 111-118. https://doi.org/10.1023/A:1012222625201.

Yamada M, A Yukita, Y Hanazumi, Y Yamahata, H Moroju, M Miyaki, T Yamasha & Histoshi S (2018). Poly(3-hydroxybutyrate) production using mannitol as a sole carbon source by Burkholderia sp. AIU M5M02 isolated from a marine environment. Journal of Fisheries science. https://doi.org/10.1007/S1256-017-1164-3

Yan D, Lu Y, Chen YF & Wu Q (2011). Waste molasses alone displaces glucose-based medium for microalgal fermentation towards cost-saving biodiesel production. Biores Tech 102(11), 6487-6493.

Yu J, Plackett D & Chen LXL (2005). Kinetics and mechanism of the monomeric products from abiotic hydrolysis of poly[(R)-3-hydroxybutyrate] under acidic and alkaline conditions. Pol Deg and Stab 89, 289-299.

Yüksekdağ ZN, Aslim B, Beyatli Y & Mercan N (2004). Effect of carbon and nitrogen sources and incubation times on poly-beta-hydroxybutyrate (PHB) synthesis by Bacillus subtilis 25 and Bacillus megaterium 12. Afr J of Biotech 3(1), 63-66

Zapata WA, Cárdenas AA & Restrep Andrés FV (2019). Evaluation of polyhydroxyalkanoate (PHAs) production with a bacterial isolate using cassava flour hydrolysates as an alternative substrate. DYNA 86(208), 75-81

Zhu C, Christopher TN, JA Perrotta A, Arthur JS & James PN (2010). Production and characterization of poly-3-hydroxybutyrate from biodiesel-glycerol by Burkholderia cepacia ATCC 17759. Biotech Progress 26(2), 424-430. doi: 10.1002/btpr.355.








How to Cite

RATNANINGRUM, D. .-., ENDAH, E. S., LISDIYANTI, P., PRIATNI, S., & SARASWATY, V. (2021). Harnessing molasses as a low-cost carbon source for production of poly-hydroxy butyrate (PHB) using Burkholderia sp. B73 bacteria. Menara Perkebunan, 89(2). https://doi.org/10.22302/iribb.jur.mp.v89i2.452

Most read articles by the same author(s)