Polyhydroxyalkanotes (PHAs) production by using canola oil as carbon source from bacteria isolated near paper pulp industry


  • Tayyaba Younas Department of Microbiology and Molecular Genetics, University of the Punjab, Quaid-i-Azam Campus, Lahore-54590, Pakistan.
  • Iftikhar Ali Department of Microbiology and Molecular Genetics, University of the Punjab, Quaid-i-Azam Campus, Lahore-54590, Pakistan.
  • Nazia Jamil Department of Microbiology and Molecular Genetics, University of the Punjab, Quaid-i-Azam Campus, Lahore-54590, Pakistan.


Samples were taken from paper pulp and mixed organic wastes of an industry for isolation of PHA producing bacterial strains. Quantitative analysis for PHA for bacterial strains was carried out by direct addition of sodium dodecyl sulphate (SDS) method. High PHA production ability was found in six strains belonging to Pseudomonas, Klebsiella, Bacillus, Streptococcus, Staphylococcus and Escherichia genera. The PHA production optimization of these six stains was done at various (NH4)2SO4 concentrations (0.2 %, 0.4 %, and 0.6%), pH (5, 6, and 7) and temperatures (4oC, 37oC, 45oC). Strain WC20 belonging to Pseudomonas sp. was found to be a potential PHA producer at 37 oC, at 0.2 % of (NH4)2SO4, using glucose as carbon source (PHA % ~28.35) and Canola Oil (PHA % ~16.06). PCR amplification of the phaC gene was also performed.


Akiyama, M., Tsuge, T. & Doi, Y. 2003. Environmental life cycle comparison of polyhydroxyalkanoates

produced from renewable carbon resources by bacterial fermentation. Polymer Degradation and

Stability 80:183-194.

Ali, I. & Jamil, N. 2014. Enhanced biosynthesis of poly (3-hydroxybutyrate) from potato starch by

Bacillus cereus strain 64-INS in a laboratory scale fermenter. Preparative Biochemistry and

Biotechnology 44: 822-833.

An derson, A. J. & Dawes, E. A.1990. Occurrence, metabolism, metabolic role, and industrial uses of

bacterial polyhydroxyalkanoates. Microbiological reviews 54:450-472.

Au subel, F. M. 2002. Short protocols in molecular biology. Wiley, New York

Cappuccino, J.G. & Sherman, N.2007. Microbiology: A Laboratory Manual. 7th ed. Pearson Education.

Casini, E., de Rijk, T. C. & de Ward, P.1997. Synthesis of poly(hydroxyalkanoate) from hydrolyzed

linseed oil. Journal of Polymer and Environment 5:153–158.

Ch audhry, W. N., Jamil, N., Ali, I., Ayaz, M. H. & Hasnain, S.2011. Screening for polyhydroxyalkanoate

(PHA)-producing bacterial strains and comparison of PHA production from various inexpensive

carbon sources. Annals of Microbiology 61:623-629.

De marco, S. M.2006. Advances in Polyhydroxyalkanoate Production in Bacteria for Biodegradable

Plastic. MMG 445 BasicBiotechnology eJournal 1:1-4.

Di n, M., Fadil, M., Ujang, Z., Van Loosdrecht, M. & Ahmad, M. A.2012. Polyhydroalkanoates (PHAs)

Production from Saponified Sunflower Oil in Mixed Cultures under Aerobic Condition. Jurnal

Teknologi 48:1–19.

Eg gink, G., Waard, P. D. & Huijberts, G. N.1995. Formation of novel poly (hydroxyalkanoates) from

long-chain fatty acids. Canadian Journal of Microbiology 41:14-21.

Es kin, N. M. & McDonald, B.1991. Canola oil. Nutrition Bulletin 16:138-146.

Fu kui, T. & Doi, Y.1998. Efficient production of polyhydroxyalkanoates from plant oils by Alcaligenes

eutrophus and its recombinant strain. Applied Microbiology and Biotechnology 49:333-336.

Ho cking, P. & Marchessault, R.1994. Biopolyesters. In: Griffin GJL (ed) Chemistry and technology of

biodegradable polymers. London, UK: Blackie Academic & Professional., pp 48-96.

Ja u, M. H., Yew, S. P., Toh, P. S., Chong, A. S., Chu, W. L., Phang, S. M., Najimudin, N. & Sudesh, K.

Biosynthesis and mobilization of poly(3-hydroxybutyrate) [P(3HB)] by Spirulina platensis.

International Journal of Biological Macromolecules 36:144-151.

Ka har, P., Tsuge, T., Taguchi, K. & Doi, Y.2004. High yield production of polyhydroxyalkanoates from

soybean oil by Ralstonia eutropha and its recombinant strain. Polymer Degradation and Stability


Ki m, M., Cho, K-S., Ryu, H. W., Lee, E. G. & Chang, Y. K.2003. Recovery of poly (3-hydroxybutyrate)

from high cell density culture of Ralstonia eutropha by direct addition of sodium dodecyl sulfate.

Biotechnology Letters 25:55-59.

Ko bayashi, G., Shiotani, T., Shima, Y. & Doi, Y.1994. Biosynthesis and Characterization of Poly

(3-hydroxybutyrate-co-3-hydroxyhexanoate) from Oils and Fats by Aeromonas sp. OL-338 and

Aeromonas sp. FA-440. In: Doi Y and Fukuda K. (eds) Biodegradable Plastics and Polymers.

Elsevier, Amsterdam, pp 410-416.

Le e, S. & Choi, J.1999. Polyhydroxyalkanoates: biodegradable polymer. In: Demain A L, Davies J E and

Atlas R M. (eds) Manual of Industrial Microbiology and Biotechnology, vol 2. 2 edn., Washington,

D.C. American Society of Microbiology, pp 616-627.

Lee, S. Y., Wong, H. H. & Choi, J. I.2000. Production of medium-chain-length polyhydroxyalkanoates

by high cell density cultivation of Pseudomonas putida under phosphorus limitation. Biotechnology

and Bioengineering 68:466–470.

Le e, S. H., Oh, D. H., Ahn, W. S., Lee, Y., Choi, Ji. & Lee, S. Y.2000. Production of poly

(3-hydroxybutyrate-co-3-hydroxyhexanoate) by high-cell-density cultivation of Aeromonas

hydrophila. Biotechnology and Bioengineering 67:240-244.

Le e, S. Y.1996. Bacterial polyhydroxyalkanoates. Biotechnology and Bioengineering 49:1-14.

Lo o, C-Y., Lee, W-H., Tsuge, T., Doi, Y. & Sudesh, K.2005. Biosynthesis and characterization of poly

(3-hydroxybutyrate-co-3-hydroxyhexanoate) from palm oil products in a Wautersia eutropha

mutant. Biotechnology Letters 27:1405-1410.

Lo o, C-Y. & Sudesh, K.2007. Polyhydroxyalkanoates: bio-based microbial plastics and their properties.

Malaysian Polymer Journal 2:31-57.

López-Cuellar, M. R., Alba-Flores, J., Rodríguez, J. N. & Pérez-Guevara, F.2011. Production of

polyhydroxyalkanoates (PHAs) with canola oil as carbon source. International Journal of Biological

Macromolecules 48: 74-80.

Ma del Rocio, L-C., Noel, G-R. J. & Fermín, P-G.2007. Production of polyhydroxyalcanoates by

Wautersia eutropha using vegetable oils as carbon source. Journal of Biotechnology 131:S156.

Mi fune, J., Nakamura, S. & Fukui, T.2008. Targeted engineering of Cupriavidus necator chromosome

for biosynthesis of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) from vegetable oil. Canadian

Journal of Chemistry 86:621-627.

Pu rushothaman, M., Anderson, R., Narayana, S. & Jayaraman, V.2001. Industrial byproducts as

cheaper medium components influencing the production of polyhydroxyalkanoates (PHA)–

biodegradable plastics. Bioprocess and Biosystems Engineering 24:131-136.

Rathinasabapathy, A., Ramsay, B. A., Ramsay, J. A. & Pérez-Guevara, F.2013. A feeding strategy for

incorporation of canola derived medium-chain-length monomers into the PHA produced by wildtype

Cupriavidusnecator. World Journal of Microbiology and Biotechnology 1-8.

Sa mbrook, J. & Russell, D. W.2001. Molecular cloning: a laboratory manual. Volume 1–3. Cold Spring

Harbor, New York: Cold Spring Harbor Laboratory Press,

Shang, L., Jiang, M., Yun, Z., Yan, H. Q. & Chang, H. N.2008. Mass production of medium-chainlength

poly (3-hydroxyalkanoates) from hydrolyzed corn oil by fed-batch culture of Pseudomonas

putida. World Journal of Microbiology and Biotechnology 24: 2783-2787.

Sp iekermann, P., Rehm, B., Kalscheuer, R. & Baumeister, D.1999. A sensitive, viable-colony staining

method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids

and other lipid storage compounds. Archives for Microbiology 171:73-80.

Tan, I. K. P., Sudesh, K. & Theanmalar, M.1997 .Saponified palm kernel oil and its major free fatty

acids as carbon substrates for the production of polyhydroxyalkanoates in Pseudomonas putida

PGA1. Applied Microbiology and Biotechnology 47:207–211.

Ts uge, T.2002. Metabolic improvements and use of inexpensive carbon sources in microbial production

of polyhydroxyalkanoates. Journal of Bioscience and Bioengineering 94:579-584.