International Journal of Pharma and Bio Sciences
    ISSN 0975-6299

Int J Pharm Bio Sci Volume 11 Issue 4, 2020 (October-December), Pages:83-95

Optimization of Parameters for Increased Production of Polyhydroxybutyrate (PHB) from Bacillus Species

Panchali Sarma, Pooja Shinde and Dr. Shampa Chakraborti *

Polyhydroxybutyrate (PHB) granules produced as a product of carbon assimilation from glucose serve as energy storage molecules in bacteria, particularly in the absence of nutrients. PHB is used to produce bioplastics due to its biodegradability. Due to the low conversion of carbon sources to PHB, optimization of the culture media, pH, temperature, etc. and utilization of promising low-cost carbon sources are essential to increase the PHB yield and reduce the cost associated with bioplastics production. The present work aimed at the maximization of PHB yield from the Bacillus spp using various alternative cheap carbon sources. A reference bacterial strain, viz., Bacillus megaterium MCC 2197(T) was procured from the National Centre for Microbial Resource, Pune. The study used Jaggery, Neera, Chikoo, Sago and Wheat bran as alternative cheap carbon sources for the production of PHB. The maximum yields of PHB obtained were 0.065 g/100 ml and 0.043 g/100 ml from Bacillus spp and the reference strain using Jaggery and Sago as carbon sources respectively with ammonium sulphate as the nitrogen source at pH 7 and a temperature of 370 C in comparison with 0.034 g/100 ml and 0.022 g/100 ml of PHB using glucose (control)  as a carbon source. The study found that Yeast Extract Glucose Medium (YEGM) and Nutrient Broth (NB) were the best media for PHB production for the Bacillus spp and Bacillus megaterium respectively. The results of this study thus confirmed that cheap alternative carbon sources under optimized conditions could significantly increase the yield of PHB from the Bacillus species.

Keywords: Bacillus species, PHB, Sudan black, cheap carbon sources.
Full HTML:
  1. Phanse N, Chincholikar A, Patel B, Rathore P, Vyas P, Patel M. Screening of PHA (poly hydroxyalkanoate) producing bacteria from diverse sources. Int. J. Biosci. 2011. 1(6):27-32.
  2. Rustagi N, Pradhan S, Singh R. Public health impact of plastics: An overview. Indian J Occup Environ Med. 2011. 15(3):100–103. Doi: 10.4103/0019-5278.93198.
  3. Pandit P, Ghate B, Kulkarni C, Mungi D, Patel T. PHB production using novel agro industrial sources from different Bacillus species. Int J Pharma Bio Sci. 2011.  2(3): 242-249.
  4. Irsath H, Santhosh S, Hemalatha V, Dhanasekar R, Dhandapani R. Production and optimization of Polyhydroxybutyrate using Bacillus subtilis BP1 isolated from Sewage samples. Int J Pure Appl Biosci. 2015.   3(1):158-166.,%20issue%201/IJPAB-2015-3-1-158-166.pdf
  5. Brandl H, Gross RA, Lenz RW, Fuller RC. Plastics from bacteria and for bacteria: poly(beta-hydroxyalkanoates) as natural, biocompatible, and biodegradable polyesters. Adv Biochem Eng Biotechnol. 1990. 41:77-93.Doi:10.1007/BFb0010232.
  6. Shah KR. Optimization and production of Polyhydroxybutyrate(PHB) by Bacillus subtilis G1S1 from soil. Int J Curr Microbiol App Sci. 2014.  3(5):377-387.
  7. Mohanrasu K, Rao R, Dinesh G, Zhang K, Prakash G, Song D et al. Optimization of media components and culture conditions for polyhydroxyalkanoates production by Bacillus megaterium  Fuel. 2020. 271(117522). Doi: 10.1016/j.fuel.2020.117522
  8. Santhi R, Balakumaran MD. Studies on utilization of fruit peel extracts for the production of polyhydroxybutyrate using Bacillus subtilis .Int J Pharm BiolSci . 2018.8(3):379-387. URL:
  9. T. L. Hartman. The Use of Sudan Black B as a Bacterial Fat Stain. Stain Technology. 1940. 15 (1) : 23-28. doi : doi:10.3109/10520294009110328
  10. Nehra K, Jaglan A, Shaheen A, Yadav J, Lathwal P, Singh M. Production of Poly-β-Hydroxybutyrate (PHB) by bacteria isolated from rhizospheric soils. Int J Microb Resour Technol. 2015.2(3):38-48.
  11. Getachew A, Woldesenbet F. Production of biodegradable plastic by polyhydroxybutyrate (PHB) accumulating bacteria using low cost agricultural waste material. BMC Res Notes. 2016. 9(1): 509-518. Doi: 10.1186/s13104-016-2321-y.
  12. Bergey, D. H. and John G Holt. Bergey's Manual of Determinative Bacteriology. Philadelphia: Lippincott Williams & Wilkins. 2000. 9th ed. 
  13. Pandit P, Ghate B, Kulkarni C, Mungi D, Patel T. PHB production using novel agro industrial sources from different Bacillus species. Int J Pharma Bio Sci. 2011.2(3):242-249.URL-
  14. Priyanka Lathwal, Kiran Nehra, Manpreet Singh, Pragati Jamdagni and Jogender S. Rana. Optimization of Culture Parameters for Maximum Polyhdroxybutyrate Production by Selected Bacterial Strains Isolated from Rhizospheric Soils. Polish Journal of Microbiology. 2015; 64 (3): 227-39. Doi: 10.5604/01.3001.0009.2118.
  15. Z. Nur Yüksekda?, Belma Asl?m, Yavuz Beyatl?, Nazime Mercan. Effect of carbon and nitrogen sources and incubation times on poly-beta-hydroxybutyrate (PHB) synthesis by Bacillus subtilis 25 and Bacillus megaterium 12. African Journal of Biotechnology. 2004. 3 (1) : 63-66. Doi: 10.5897/AJB2004.000-2011
  16. Yüksekda? Z N, Asl?m B, Beyatl? Y, Mercan N.Effect of carbon and nitrogen sources and incubationtimes on poly-beta-hydroxybutyrate (PHB) synthesis byBacillus subtilis 25 and Bacillus megaterium 12. Afr. J. Biotechnol. 2004.3(1):63-66. Doi: 10.5897/AJB2004.000-2011.
  17. Salman Ahmady-Asbchin, Hassan Rezaee, Moein Safari, Pantea Zamanifar, Davood Siyamiyan. Production and optimization of polyhydroxybutyrate (PHB) from Bacillus megaterium as biodegradable plastic. European Journalof Biological Research. 2020. 10(1): 26-34. Doi: 10.5281/zenodo.3711400.
  18. Manju J , Prabakaran P. Effect of carbon sources in the production of polyhydroxybutyrate(PHB) by Bradyrhizobium and Rhizobium sp.from Aeschynomene indica. International Journal of Research and Analytical Reviews. 2019; 6(2):823-827.
  19. Nur Arfa Yanti, Langkah Sembiring, Sebastian Margino. Production of Poly-α-hydroxybutyrate (PHB) from Sago Starch by The Native Isolate Bacillus megaterium PSA10. Indonesian Journal of Biotechnology. 2009;14(1):1111-16. Doi: 10.22146/ijbiotech.7559.
  20. G Sathiyanarayanan, G saibaba, G Seghal Kiran, Joseph Selvin. Process optimization and production of polyhydroxybutyrate using palm jaggery as economical carbon source by marine-sponge associated Bacillus licheniformis MSBN12. Bioprocess Biosyst Eng. 2013.  36(12):1817-27. Doi: 10.1007/s00449-013-0956-9.
  21. John H. Law, Ralph A. Slepecky. Assay of Poly-β-Hydroxybutyric acid. J Bacteriol. 1961. 82(1): 33–36. URL:
  22. Full TD, Jung MT, Madigan O. Production of polyhydroxyalkanoates from soy molasses oligosaccharides by new, rapidly growing Bacillus species. Lett Appl Microbiol. 2006.43(4):377–84. Doi: 10.1111/j.1472-765X.2006.01981.x.
  23. Vishnuvardhan Reddy S, Thirumala M, Mahmood S. Production of PHB and P (3HB-co-3HV) biopolymers by Bacillus megaterium strain OU303A isolated from municipal sewage sludge. World J Microbiol Biotechnol. 2009.25:391–397. doi: 10.1007/s11274-008-9903-3.
  24. Fatimah Alshehrei. Production of Polyhydroxybutyrate (PHB) by Bacteria Isolated from Soil of Saudi Arabia. Journal of Pure Applied Microbiology. 2019. 13 (2): 897-904. doi: 10.22207/JPAM.13.2.26 
  25. Seid Mohammed,Himadri Tanaya Behera,Aman Dekebo,Lopamudra Ray.Optimization of the culture conditions for production of Polyhydroxyalkanoate and its characterization from a new Bacillus cereus sp. BNPI-92 strain, isolated from plastic waste dumping yard. International Journal of Biological Macromolecules. 2019. doi: 10.1016/j.ijbiomac.2019.11.138.
  26. E. Gayathiri, B. Bharathi, N. Siva, R. Prabavathi and S.Velu. Production, Optimization and Characterization of
    Polyhydroxybutryate by Bacillus subtilis Isolated from Garden Soil. International Journal of Pharma And Chemical Research. 2017. 3(2): 155 - 165.
  27. Christina Thapa, Pallavi Shakya, Rabina Shrestha, Sushovita Pal, Prakash Manandhar. Isolation of Polyhydroxybutyrate (PHB) Producing Bacteria, Optimization of Culture Conditions for PHB production, Extraction and Characterization of PHB. Nepal Journal of Biotechnology. 2019. 6(1), 62-68. doi: 10.3126/njb.v6i1.22339
  28. G Sathiyanarayanan, G Seghal Kiran, Joseph Selvin, G Saibaba. Optimization of polyhydroxybutyrate production by marine Bacillus megaterium MSBN04 under solid state culture. Int J Biol Macromol. 2013. 60: 253-61.doi:10.1016/j.ijbiomac.2013.05.031.
  29. Priya Kumari and Harish Kumar Dhingra. Isolation and Characterization of PHB producing microogranisms isolated from root nodules of leguminous plants. The Bioscan. 2013. 8(1): 109-113.
  30. Nur Arfa Yanti, Langkah Sembiring, Sebastian Margino, Nurhayani. H. Muhiddin. A study on production of Poly-β-Hydroxybutyrate bioplastic from sago starch by indigenous amylolytic bacteria. Indonesian Journal of Biotechnology. 2013. 18 (2): 144-150. doi: 10.22146/IJBIOTECH.7877.
  31. Mascarenhas Joyline and Aruna K. Production and characterization of polyhydroxyalkanoates (PHA) by bacillus megaterium strain JHA using inexpensive agro-industrial wastes. International Journal of Recent Scientific Research. 2019. 10 ( 7 ) : 33359-33374. doi: 10.24327/ijrsr.2019.1007.3656
  32. Suresh S. Suryawanshi,Sushant S. Sarje,Prakash C. Loni, Sourabh bhujbal , Prajakta P. Kamble. Bioconversion of Sugarcane Molasses into Bioplastic (Polyhydroxybutyrate) using Bacillus cereus 2156 under Statistically Optimized Culture Conditions. Analytical Chemistry Letters. 2020. 10(1):80-92. doi: 10.1080/22297928.2020.1746197.
  33. Watsana Penkhrue, Dieter Jendrossek, Chartchai Khanongnuch, Wasu Pathom-aree, Tomoyasu Aizawa, Rachel L. Behrens, S. Lumyong. Response surface method for polyhydroxybutyrate (PHB) bioplastic accumulation in Bacillus drentensis BP17 using pineapple peel. Plos One. 2020. 15(3):1-21. e0230443. doi:10.1371/journal.pone.0230443.
  34. Abdullah A. Alarfaj, Mohammed Arshad, Essam N. Sholkamy, Murugan A Munusamy. Extraction and Characterization of Polyhydroxybutyrates (PHB) from Bacillus thuringiensis KSADL127 isolated from mangrove environments of Saudi Arabia. Braz. arch. biol. technol. 2015; 58(5):781-788. doi: 10.1590/S1516-891320150500003.
  35. Salman Ahmad, Abuzer Amir , Md. Zafaryab , Khwaja Osama , Soban Ahmad Faridi , Mohammed Haris Siddiqui , Moshahid A Rizvi M and Md. Asad Khan. Production and Characterization of Polyhydroxybutryrate Biopolymer from Azohydromonas australica Using Sucrose as a Sole Carbon Source. J Microb Biochem Technol. 2017.  9 (3):82-86. doi:10.4172/1948-5948.1000348.
[Download PDF]
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy
Pharmaceutical Fields
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Pharmaceutics
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Novel drug delivery system
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Nanotechnology
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Pharmacology
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Pharmacognosy
© Copyright 2009-2015 IJPBS, India. All rights reserved. Specialized online journals by ubijournal. Website by Ubitech Solutions
         Home I Contact I Terms & Conditions