International E-publication: Publish Projects, Dissertation, Theses, Books, Souvenir, Conference Proceeding with ISBN.  International E-Bulletin: Information/News regarding: Academics and Research

Production of Polyhydroxybutyrate (phb) by Pseudomonas Putida Strain kt2440 on Cassava Hydrolysate Medium

Author Affiliations

  • 1Department of Chemical Engineering, Ladoke Akintola University of Technology, Ogbomoso, NIGERIA
  • 2Department of Chemical Engineering, Obafemi Awolowo University, Ile-Ife, NIGERIA
  • 3National Biotechnology Development Agency (NABDA), Abuja, NIGERIA

Res.J.chem.sci., Volume 1, Issue (4), Pages 67-73, July,18 (2011)


The potential of local strain of Pseudomonas putida strain KT2440 in polyhydroxbutyrate production was investigated in this study. This was done to establish the capabilities of local strains to utilize renewable and locally available substrates in polyhydroxybutyrate production. It involved hydrolysis of extracted starch from freshly harvested cassava tubers using enzyme-enzyme hydrolysis method, followed by aerobic fermentation using Pseudomonas putida in batch cultures on a mixture of the hydrolysate and nutrient media. The reducing sugar hydrolysate served as the carbon source while di- ammonium sulphate was the source of Nitrogen. The reaction temperature, pH and agitation rate in the fermentor were maintained at 30 C, 7.5 and 400 rpm respectively. The biomass growth was measured by cell dry weight and the polyhydroxybutyrate content measured by gas chromatographic method. The results obtained showed that the medium supported the growth of the organism. After 72 h fermentation, the substrate consumption by the organism was 8.88 g/l to give a dry cell weight of 0.91 g/l, resulting in a biomass yield on substrate (Yx/s) of 0.1025 g g-1. The gas chromatographic analysis gave a final polyhydroxybutyrate value of 0.2285 g/l with corresponding product yield on biomass (Y p/x) of 0.2511 g g-1 (25.11%) and product yield on substrate (Yp/s) of 0.0257 g g-1. It can be concluded that Pseudomonas putida strain KT2440 has capability to utilize cheap, renewable and locally available substrates in polyhydroxybutyrate production. Also, comparing the result of the present study with those from the previous ones showed that although Pseudomonas putida strain KT2440 accumulated polyhydroxybutyrate lower than the usual value in excess of 50 % of the cell dry weight given by various strains of microorganisms during polyhydroxybutyrate production it is a potential candidate for polyhydroxybutyrate production.


  1. Anderson A. J. and Dawes E. A., Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates, Microbiol. Rev., 54, 450-472 (1990)
  2. Aransiola E.F., Betiku E., Adetunji O.A. and Solomon B.O., Production of Baker’s Yeast Saccharomyces cerevisiae) from raw Cassava Starch Hydrolysates in a Bioreactor under Batch Process. Biotechnol.,5(1), 98 – 103 (2006)
  3. Aransiola E.F., Adetunji O.A., Aremu M.O. and Solomon B.O., Fed-batch production of Baker’s yeast (Saccharomyces Cerevisiae) from cassava Manihot esculenta) starch hydrolysate, Ife J. Sci., 10(1), 39 – 44 (2008)
  4. Aremu M.O., Layokun S.K. and Solomon B.O., Production of poly (3- hydroxybutyrate) from cassava starch hydrolysate by Pseudomonas aeruginosa NCIB 950. Am. J. Sci. Ind. Res., 1(3), 421-426 (2010a)
  5. Beaulieu M., Beaulieu Y., Melinard J., Pandian S. and Goulet J., Influence of ammonium salts and cane molasses on growth of Alcaligenes eutrophus and production of polyhydroxybutyrate, Appl, Environ, Microbiol, 61,165-168 (1995)
  6. Brandl H., Gross R. A., Lenz R.W. and Fuller R.C., Pseudomonas oleovorans as a source of poly (-hydroxyalkanoates) for potential applications as biodegradable polyesters. Appl. Environ. Microbiol.54, 1977-1982(1988)
  7. Choi J. and Lee S.Y., Factors affecting the economics of polyhydroxyalkaonoate production by bacterial fermentation. Appl. Microbiol. Biotechnol. 51, 13-21(2001a)
  8. Du G., Si Y., and Yu J, Inhibitory effect of medium-chain-length fatty acid on synthesis of polyhydroxyalkanoates from volatile fatty acid by Ralstonia eutrophus.Biotechnol. Lett.23, 613-617 (2001a)
  9. Du G. and Yu J, Green technology for conversion of food scraps to biodegradable thermoplastic polyhydroxyalkanoates. Environ. Sci.Technol.36, 511 – 516 (2002b)
  10. El-Holi M. A. and Al-Delaimy K.S., Citric Acid Production from Whey with Sugars and Additives by Aspergillus niger. Afr. J. Biotechnol.2, 356-359 (2003)
  11. Gaouar O., Aymard C., Zakhia N. and Rios G.M., Enzymatic hydrolysis of cassava starch into maltose syrup in a continuous membrane reactor, J. Chem. Technol. Biotechnol.,69, 367–375 (1997b)
  12. Hocking P.J. and Marchessault R.H., Biopolyesters. In Chemistry and Technology of Biodegradable Polymers ed. Griffin, G.M.L. Blackie Academic and Professional, Chapman & Hall., 48-96 (1994)
  13. Kim B.S., Production of poly (3-hydroxybutyrate) from inexpensive substrates, Enzy. Microb. Technol.,27, 774-777 (2000)
  14. Leaversuch R. D., Industry weighs need to make polymers biodegradable, Mod. Plast. 64, 52-55 (1987)
  15. Lee B., Pometto A. L., Fratzke A. and Bailey T. B., Biodegradation of Degradable plastic polythene by Phanerochaeteand Streptomyces speciesAppl. Environ. Microbiol. 57, 678-685(1991)
  16. Lee S.Y., Bacterial Polyhydroxyalkanoates, A Review. Biotechnol. Bioeng. 49,1-14 (1996)
  17. Marchessault, P., Fermentation methods for theproduction of poly (3-hydroxybutyrate) by Alcaligeneseutrophus DSM 545. MSc Thesis,McGill University Canada (1996)
  18. Miller G.L., Use of dinitrosalicylic acid reagent for determination of reducing sugar, Anal. Chem.,31, 426-428 (1959)
  19. Omemu A.M., Akpan I., Bankole M.O. and Teniola O.D., Hydrolysis of raw tuber starches by amylase of Aspergillus niger AM07 isolated from the soil. Afr. J. Biotechnol.4, 19–25(2005)
  20. Poirier Y., Nawrath C. and Somerville C., Production of polyhydroxyalkanoate, A family of Biodegradable plastics and Elastomers in bacteria and plants, A review, Biotechnol.,13, 142-150 (1995)
  21. Purushothaman M.A., Narayama R.K.I.S., Jayaraman V.K., Industrial byproducts as cheaper medium components inuencing the production of polyhydroxyalkanoates (PHA)-biodegradable plastics, Bioproc. Biosys. Engineering24, 131–136 (2001)
  22. Riis V. and Mai I., Gas chromatographic determination of poly--hydroxybutyric acid in microbial biomass after hydrochloric acid propanolysis. J. Chromatograp.445, 285-9(1988)
  23. Rusendi D. and Sheppard J.D., Hydrolysis of potato processing waste for the production of poly--hydroxybutyrate, Elsevier J. Biores. Tech.,54, 191-196 (1995)
  24. Santimano M.C., Nimali N. P. and Garq S., PHA Production Using Low-Cost Agro- Industrial Wastes by Bacillus sp. Strain COL1/A6. J. Microbiol, 4(3), 89 – 96 (2009)
  25. Steinbüchel A. and Fuchtenbusch B., Bacteria and other biological systems for polyester production. TIBTEC,16, 419-427 (1998)
  26. Suzuki T., Yamane T. and Shimizu S., Mass Production of Poly-- hydroxybutyric Acid by Fully Automatic Fed-batch Culture of Methylotroph. Appl, Microbiol, Biotechnol,23, 322-329 (2001)
  27. Wu Q., Huang H., Hu G.H., Chen J., Ho K.P. and Chen G.Q., Production of poly- 3- hydroxybutyrate by Bacillus sp. JMa5 cultivated in molasses media. Antonie Van Leeuwenhoek 80, 111-118 (2001)
  28. Yasser E. and Alexander S., Large - scale production of Poly (3-hydroxyoctanoic acid) by Pseudomonas Putida GP01 and a simplified downstream process. Appl. Environ. Microbiol.75(3), 643-651(2009)