Research Article | Volume: 5, Issue: 5, Sep-Oct, 2017

Fermentative Production of Polyhydroxyalkanoates (PHAs) from Glycerol by Zobellella taiwanensis Azu-IN1

Mohamed Ali Abdel-Rahman Said El-Sayed Desouky Mohamed Salah Azab Mahmoud E. Esmael   

Open Access   

Published:  Oct 30, 2017

DOI: 10.7324/JABB.2017.50503

Polyhydroxyalkalonates (PHAs) are biodegradable thermoplastics that are receiving immense attention as an alternative to petroleum derived plastics. The present study aimed to produce PHA using cheap and ecofriendly biodiesel waste glycerol as a substrate. Several PHA–producing bacterial isolates were isolated from different environmental samples and their efficacy for PHA production was assessed. Isolate Azu-IN1 showed the highest PHA production and was identified as Zobellella taiwanensis Azu-IN1 using 16S rRNA gene sequence and biochemical characterization. Factors affecting PHA production were optimized in batch fermentations. Supplementation of ammonium chloride as nitrogen source, methanol as an auxiliary carbon source, and agitation rate are the main limiting factors affecting PHA production. Maximum PHA production of 2.65 g/L at recovery yield of 50.3 (%), w/w) was achieved after 36 h in batch fermentation using optimized medium. Enhanced PHA production of 3.73 g/L with recovery yield of 61.7 % (w/w) was obtained in fed batch fermentation. The characteristics of extracted PHA were analyzed using Fourier Transform Infrared Spectroscopy (FTIR) 1H and 13C Nuclear Magnetic Resonance (NMR) spectroscopy. This is the first report on accumulation of PHA by Zobellella taiwanensis using glycerol as the sole carbon source.

Keyword:     Bioplastic; PHA-production; fed-batch fermentationglycerol; Zobellella taiwanensis.


Abdel-Rahman MA, Desouky SE, Azab MS, Esmael ME. Fermentative Production of Polyhydroxyalkanoates (PHAs) from Glycerol by Zobellella taiwanensis Azu-IN1. J App Biol Biotech. 2017; 5 (05): 16-25.

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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1. Wong SL, Ngadi N, Abdullah TAT, Inuwa IM. Current state and future prospects of plastic waste as source of fuel: A review. Renewable and Sustainable Energy Reviews. 2015; 50: 1167-1180.

2. Ray S, Prajapati V, Patel K, Trivedi U. Optimization and characterization of PHA from isolate Pannonibacter phragmitetus ERC8 using glycerol waste. International journal of biological macromolecules. 2016; 86: 741-749.

3. Salehizadeh H, Loosdrecht MCMV. Production of polyhydroxyalkanoates by mixed culture: recent trends and biotechnological importance. Biotechnology Advances. 2004; 22: 261-279.

4. Castilho LR, Mitchell DA, Freire DMG. Production of polyhydroxyalkanoates from waste materials and by-products by submerged and solid-state fermentation. Bioresource Technology. 2009; 100(23): 5996-6009.

5. Bugnicourt E, Cinelli P, Lazzeri A, Alvarez V. Polyhydroxyalkanoate: Review of synthesis, characteristics, processing and potential applications in packaging. Express Polymer Letters. 2014; 8 (11): 791-808.

6. Muhammadi S, Muhammad A, Shafqat, H. Bacterial polyhydroxyalkanoates-eco-friendly next generation plastic: Production, biocompatibility, biodegradation, physical properties and applications. Green Chemistry Letters and Reviews. 2015; 8(3-4): 56-77.

7. Anjum A, Zuber M, Mahmood, ZK, Noreen, A, Naveed A and Tabasum, S. Microbial production of polyhydroxyalkanoates and its copolymers. International Journal of Biological Macromolecules. 2016; 89:161-174.

8. Ahn WS, Park SJ, Lee SY. Production of poly (3-hydroxybutyrate) from whey by cell recycle fed-batch culture of recombinant Escherichia coli. Biotechnology Letter. 2001; 23: 235-240.

9. da Silva GP, Mack M, Contiero J. Glycerol: A promising and abundant carbon source for industrial microbiology. Biotechnology Advances. 2009; 27(1): 30-9.

10. Du C, Sabirova J, Soetaert W, Lin SKC. Polyhydroxyalkanoates production from low-cost sustainable raw materials. Current Chemical Biology. 2012; 6(1): 14-25.

11. Spiekermann P, Rehm B, Kalscheuer R. A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds. Archives of Microbiology. 1999; 71: 73-80.

12. Abdel-Rahman MA. Establishment of efficient cellulolytic bacterial consortium potential for designed composting of rice straw. International Journal of Advanced Research in Biological Science. 2016; 3(4): 211-228.

13. Kuhn J, Müller H, Salzig D, Czermak P. A rapid method for an offline glycerol determination during microbial fermentation. Electronic Journal of Biotechnology. 2015; 18(3): 252-255.

14. Sathianachiyar S, Devaraj A. Biopolymer production by bacterial species using glycerol, aby product of biodiesel. International Journal of Scientific and Research Publications. 2013; 3(8): 1-5.

15. Silverstein R, Bassler G, Morrill T. Spectrometric identification of organic compounds, 5th ed. John Wiley & Sons, INC, New York. 1991

16. Gorenflo V, Steinbüchel A, Marose S. Quantification of bacterial polyhydroxy-alkanoic acids by Nile red staining. Applied Microbiolology and Biotechnology. 1999; 51:765-772.

17. Ibrahim MHA, Steinbüche A. Zobellella denitrificansstrain MW1. a newly isolated bacterium suitable for poly (3hydroxybutyrate) production from glycerol. Journal of Applied Microbiology. 2010; 108(1): 214-225.

18. El-Abd MA, El-Sheikh HH, Desouky S, Shehab A. Identification, Biodegradation and bio-evaluation of biopolymer produced from Bacillus thuringenesis. Journal of Applied Pharmaceutical Science. 2017; 7 (04): 103-110.

19. Sindhu R, Ammu B, Binod P, Deepthi SK, Ramachandran, K B, Soccol, C R, Pandey, A. Production and characterization of poly-3hydroxybutrate from crude glycerol by Bacillus spharaeicus NII 0838 and improving its thermal properties by blending with other polymers. Brazilian archives of Biology and Technology. 2011; 54(4): 783-794.

20. Chanasit W, Hodgson B, Sudesh K, Umsakul K. Efficient production of polyhydroxyalkanoates from Pseudomonas mendocina PSU using a biodiesel liquid waste (BLW) as the sole carbon source. Bioscience, biotechnology, and biochemistry. 2016; 80(7): 1440-1450.

21. de Castro JS, Nguyen LD, Seppala J. Bioconversion of commercial and waste glycerol into value-added polyhydroxyalkanoates by bacterial strains. Journal of Microbial & Biochemical Technology. 2014; 6(6): 337-345.

22. de Paula FC, Kakazu S, de Paula CBC, Gomez JGC, Contiero J. Polyhydroxyalkanoate production from crude glycerol by newly isolated Pandoraea sp. Journal of King Saud University-Science. 2017; 29(2), 166-173.

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

24. Aramvash A, Akbari SZ, Dashti AS, Ghafari, MD. Statistical physical and nutrient optimization of bioplastic polyhydroxy butyrate production by Cupriavidus necator. International Journal of Environmental Science and Technology. 2015; 12(7):2307-2316.

25. Dashti AS, Aramvash A. One-Factor-at-a-Time Optimization of Polyhydroxybutyrate Production and Growth of Alcaligenes eutrophus by Altering Culture Parameters and Incubation Time. Journal of Sciences. 2015; 26(4): 303-313.

26. Sasidharan RS, Bhat, SG, Chandrasekaran M. Biocompatible polyhydroxybutyrate production by marine Vibrio azureus BTKB33 under submerged fermentation. Annals of microbiology. 2015; 65 (1): 455-465.

27. Kalaiyezhini D, Ramachandran KB. Biosynthesis of poly-3- hydroxybutyrate from glycerol by Paracoccus denitrificansin a batch bioreactor: Effect of process variables. Preparative Biochemistry and Biotechnology. 2015; 45(1): 69-83.

28. Ramadas NV, Singh SK, Socool C R, Pande A. Polyhydroxylbutrate production using agro-industrial residue as substrate by Bacillus sphaericus NCIM 5149. Brazilian archives of Biology and Technology. 2009; 52(1): 17-23.

29. Sharma P, Bajaj BK. Cost-effective substrates for production of-poly-β-hydroxybutrate by a newly isolated Bacillus cereus PS-10. Journal of environmental biology. 2015; 36: 1297-1304.

30. Tripathi AD, Yadav A, Jha A, Srivastava SK. Utilizing of sugar refinery waste (Cane Molasses) for production of bio-plastic under submerged fermentation process. Journal of Polymers and the Environment. 2012; 20: 446-453.

31. Serafim LS, Lemos PC, Albuquerque MGE, Reis MAM. Strategies for PHA production by mixed cultures and renewable waste materials. Applied microbiology and biotechnology. 2008; 81(4), 615-628.

32. Wei YH, Chen WC, Huang CK, Wu HS, Sun YM, Lo CW. Screening and evaluation of polyhydroxybutyrate-producing strains from indigenous isolate Cupriavidus denitrificans strains. International journal of molecular sciences. 2011; 12(1): 252-265.

33. Braunegg G, Genser K, Bona R, Haage G, Schellauf F, Winkler E. Production of PHAs from agricultural waste material. In: Macromolecular Symposia (Vol 144, No 1, pp 375-383) WILEY‐VCH Verlag GmbH & Co KGaA. 1999.

34. Dobroth ZT, Hu S, Coats ER, McDonald AG. Polyhydroxybutyrate synthesis on biodiesel wastewater using mixed microbial consortia. Bioresource Technology. 2011; 102(3): 3352-3359.

35. Yezza A, Fournier D, Halasz A, Hawari J. Production of polyhydroxyalkanoates from methanol by a new methylotrophic bacterium Methylobacterium sp GW2. Applied Microbiology and Biotechnology. 2006; 73: 211-218.

36. Zhu C, Nomura CT, Perrotta JA, Stipanovic AJ, Nakas AP. Production and characterization of poly-3-hydroxybutyrate from biodiesel-glycerol by Burkholderia cepacia ATCC 17759. Biotechnology progress. 2010; 26(2):424-430.

37. Cui B, Huang S, Xu F, Zhang R, Zhang Y. Improved productivity of poly (3-hydro xybutyrate) (PHB) In thermophilic Chelatococcus daeguensis TAD1 using glycerol as the growth substrate in a fed-batch culture. Applied microbiology and biotechnology. 2015; 99(14):6009-6019.

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