Skip to main content
SpringerLink
Log in

Enhancement of bioplastic polyhydroxybutyrate P(3HB) production from glucose by newly engineered strain Cupriavidus necator NSDG-GG using response surface methodology

  • Original Article
  • Published:
3 Biotech Aims and scope Submit manuscript

Abstract

This study aimed to enhance production of polyhydroxybutyrate P(3HB) by a newly engineered strain of Cupriavidus necator NSDG-GG by applying response surface methodology (RSM). From initial experiment of one-factor-at-a-time (OFAT), glucose and urea were found to be the most significant substrates as carbon and nitrogen sources, respectively, for the production of P(3HB). OFAT experiment results showed that the maximum biomass, P(3HB) content, and P(3HB) concentration of 8.95 g/L, 76 wt%, and 6.80 g/L were achieved at 25 g/L glucose and 0.54 g/L urea with an agitation rate of 200 rpm at 30 °C after 48 h. In this study, RSM was applied to optimize the three key variables (glucose concentration, urea concentration, and agitation speed) at a time to obtain optimal conditions in a multivariable system. Fermentation experiments were conducted in shaking flask by cultivation of C. necator NSDG-GG using various glucose concentrations (10–50 g/L), urea concentrations (0.27–0.73 g/L), and agitation speeds (150–250 rpm). The interaction between the variables studied was analyzed by ANOVA analysis. The RSM results indicated that the optimum cultivation conditions were 37.70 g/L glucose, 0.73 g/L urea, and 200 rpm agitation speed. The validation experiments under optimum conditions produced the highest biomass of 12.84 g/L, P(3HB) content of 92.16 wt%, and P(3HB) concentration of 11.83 g/L. RSM was found to be an efficient method in enhancing the production of biomass, P(3HB) content, and P(3HB) concentration by 43, 21, and 74%, respectively.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Almeida A, Giordano AM, Nikel PI, Pettinari MJ (2010) Effects of aeration on the synthesis of poly(3-hydroxybutyrate) from glycerol and glucose in recombinant Escherichia coli. Appl Environ Microbiol 76:2036–2040

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Anderson AJ, Dawes WA (1990) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev 54:450–472

    CAS  PubMed  PubMed Central  Google Scholar 

  • Aramvash A, Shahabi ZA, Aghjeh SD, Ghafari MD (2015) Statistical physical and nutrient optimization of bioplastic polyhydroxybutyrate production by Cupriavidus necator. Int J Environ Sci Technol 12:2307–2316

    Article  CAS  Google Scholar 

  • Bezerra MA, Santelli RE, Oliveira EP, Villar LS, Escaleira LA (2008) Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta 76:965–977

    Article  CAS  PubMed  Google Scholar 

  • Braunegg G, Sonnleitner BY, Lafferty RM (1978) A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass. Eur J Appl Microbiol Biotechnol 6:29–37

    Article  CAS  Google Scholar 

  • Budde CF, Mahan AE, Lu J, Rha C, Sinskey AJ (2010) Roles of multiple acetoacetyl coenzyme a reductases in polyhydroxybutyrate biosynthesis in Ralstonia eutropha H16. J Bacteriol 192:5319–5328

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Daneshi A, Younesi H, Ghasempouri SM, Sharifzadeh M (2010) Production of poly-3-hydroxybutyrate by Cupriavidus necator from corn syrup: statistical modeling and optimization of biomass yield and volumetric productivity. J Chem Technol Biotechnol 85:1528–1539

    CAS  Google Scholar 

  • Dashti MG, Abdeshahian P, Yusoff WMW, Kalil MS, Hamid AA (2014) Repeated batch fermentation biotechnology for the biosynthesis of lipid and gamma-linolenic acid by Cunninghamella bainieri 2A1. Biomed Res Int. https://doi.org/10.1155/2014/831783

    Article  Google Scholar 

  • Dashti MG, Abdeshahian P, Sudesh K, Phua KK (2016) Optimization of Salmonella Typhi biofilm assay on polypropylene microtitre plates using response surface methodology. Biofouling 32:477–487

    Article  CAS  Google Scholar 

  • Dawes EA, Senior PJ (1973) The role and regulation of energy reserve polymers in micro-organisms. In: Rose AH, Tempest DW (eds) Advances in microbial physiology, 11th edn. Academic Press, Cambridge, pp 135–266

    Google Scholar 

  • Emeko HA, Olugbogi AO, Betiku E (2015) Appraisal of artificial neural network and response surface methodology in modeling and process variable optimization of oxalic acid production from cashew apple juice: a case of surface fermentation. Bioresources 10:2067–2082

    Article  Google Scholar 

  • Getachew A, Woldesenbet F (2016) Production of biodegradable plastic by polyhydroxybutyrate (PHB) accumulating bacteria using low cost agricultural waste material. BMC Res Notes 9:509

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hembree B, Prins J, Spagon P, Tobias P, Zey C (2010) NIST/SEMATECH engineering statistics handbook. https://www.itl.nist.gov/div898/handbook. Accessed 17 Sept 2010

  • Jiang G, Hill DJ, Kowalczuk M, Johnston B, Adamus G, Irorere V, Radecka I (2016) Carbon sources for polyhydroxyalkanoates and an integrated biorefinery. Int J Mol Sci 17:1157

    Article  CAS  PubMed Central  Google Scholar 

  • Kadier A, Abdeshahian P, Kalil MS, Hamid AA (2018) Optimization of the key medium components and culture conditions for efficient cultivation of G. sulfurreducens strain PCA ATCC 51573 using response surface methodology. Iran J Sci Technol Trans A Sci 42:237–244

    Article  Google Scholar 

  • Kamble AL, Meena VS, Banerjee UC (2010) Effect of agitation and aeration on the production of nitrile hydratase by Rhodococcus erythropolis MTCC 1526 in a stirred tank reactor. Lett Appl Microbiol 51:413–420

    Article  CAS  PubMed  Google Scholar 

  • Lee SY (1996) Plastic bacteria? Progress and prospects for polyhydroxyalkanoate production in bacteria. Trends Biotechnol 14:431–438

    Article  CAS  Google Scholar 

  • Mifune J, Nakamura S, Fukui T (2008) Targeted engineering of Cupriavidus necator chromosome for biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from vegetable oil. Can J Chem 86:621–627

    Article  Google Scholar 

  • Mokhtari-Hosseini ZB, Vasheghani-Farahani E, Heidarzadeh-Vazifekhoran A, Shojaosadati SA, Karimzadeh R, Darani KK (2009) Statistical media optimization for growth and PHB production from methanol by a methylotrophic bacterium. Bioresour Technol 100:2436–2443

    Article  CAS  PubMed  Google Scholar 

  • Montgomery CD (1991) Design and analysis of experiments. Wiley, New York

    Google Scholar 

  • Narayanan A, Ramana KV (2012) Polyhydroxybutyrate production in Bacillus mycoides DFC1 using response surface optimization for physico-chemical process parameters. 3 Biotech 2:287–296

    Article  PubMed Central  Google Scholar 

  • Orita I, Iwazawa R, Nakamura S, Fukui T (2012) Identification of mutation points in Cupriavidus necator NCIMB 11599 and genetic reconstitution of glucose-utilization ability in wild strain H16 for polyhydroxyalkanoate production. J Biosci Bioeng 113:63–69

    Article  CAS  PubMed  Google Scholar 

  • Pandian SR, Deepak V, Kalishwaralal K, Rameshkumar N, Jeyaraj M, Gurunathan S (2010) Optimization and fed-batch production of PHB utilizing dairy waste and sea water as nutrient sources by Bacillus megaterium SRKP-3. Bioresour Technol 101:705–711

    Article  CAS  PubMed  Google Scholar 

  • Poirier Y, Dennis DE, Klomparens K, Somerville C (1992) Polyhydroxybutyrate, a biodegradable thermoplastic, produced in transgenic plants. Science 256:520–523

    Article  CAS  PubMed  Google Scholar 

  • Raberg M, Kaddor C, Kusian B, Stahlhut G, Budinova R, Kolev N, Bowien B, Steinbüchel A (2012) Impact of each individual component of the mutated PTSNag on glucose uptake and phosphorylation in Ralstonia eutropha G+1. Appl Microbiol Biotechnol 95:735–744

    Article  CAS  PubMed  Google Scholar 

  • Sangkharak K, Prasertsan P (2007) Optimization of polyhydroxybutyrate production from a wild type and two mutant strains of Rhodobacter sphaeroides using statistical method. J Biotechnol 132:331–340

    Article  CAS  PubMed  Google Scholar 

  • Sathiyanarayanan G, Saibaba G, Kiran GS, Selvin J (2013) A statistical approach for optimization of polyhydroxybutyrate production by marine Bacillus subtilis MSBN17. Int J Biol Macromol 59:170–177

    Article  CAS  PubMed  Google Scholar 

  • Shah AA, Hasan F, Hameed A, Ahmed S (2008) Biological degradation of plastics: a comprehensive review. Biotechnol Adv 26:246–265

    Article  CAS  PubMed  Google Scholar 

  • Sharma L, Singh AK, Panda B, Mallick N (2007) Process optimization for poly-β-hydroxybutyrate production in a nitrogen fixing cyanobacterium, Nostoc muscorum using response surface methodology. Bioresour Technol 98:987–993

    Article  CAS  PubMed  Google Scholar 

  • Sheu DS, Chen WM, Yang JY, Chang RC (2009) Thermophilic bacterium Caldimonas taiwanensis produces poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from starch and valerate as carbon sources. Enzyme Microb Technol 44:289–294

    Article  CAS  Google Scholar 

  • Sudesh K (2013) Polyhydroxyalkanoates from palm oil: biodegradable plastics. Springer Science and Business Media, Heidelberg

    Book  Google Scholar 

  • Surwase SN, Jadhav SB, Phugare SS, Jadhav JP (2013) Optimization of melanin production by Brevundimonas sp. SGJ using response surface methodology. 3 Biotech 3:187–194

    Article  PubMed  Google Scholar 

  • Tripathi AD, Srivastava SK, Singh RP (2013) Statistical optimization of physical process variables for bio-plastic (PHB) production by Alcaligenes sp. Biomass Bioenergy 55:243–250

    Article  CAS  Google Scholar 

  • Wang J, Wan W (2009) Experimental design methods for fermentative hydrogen production: a review. Int J Hydrog Energy 34:235–244

    Article  CAS  Google Scholar 

  • Wisuthiphaet N, Napathorn SC (2016) Optimisation of the use of products from the cane sugar industry for poly(3-hydroxybutyrate) production by Azohydromonas lata DSM 1123 in fed-batch cultivation. Process Biochem 51:352–361

    Article  CAS  Google Scholar 

  • Yu L, Lei T, Ren X, Pei X, Feng Y (2008) Response surface optimization of l-(+)-lactic acid production using corn steep liquor as an alternative nitrogen source by Lactobacillus rhamnosus CGMCC 1466. Biochem Eng J 39:496–502

    Article  CAS  Google Scholar 

  • Zafar M, Kumar S, Kumar S, Dhiman AK (2012) Optimization of polyhydroxybutyrate (PHB) production by Azohydromonas lata MTCC 2311 by using genetic algorithm based on artificial neural network and response surface methodology. Biocatal Agric Biotechnol 1:70–79

    CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by Research University Grant (RUI) from Universiti Sains Malaysia (1001/PBIOLOGI/811328). Also, this study contributed to the international research project PHABIO APP – Polyhydroxyalkanoate Biopolymers from Animal Waste Fats for the Production of Value Added Biobased and Biodegradable Bioplastic Materials, founded by the Federal Ministry of Education and Research of Germany and supervised by the PTJ Jülich. Nazila Biglari expresses her in-depth gratitude towards Universiti Sains Malaysia for supporting her doctoral studies through USM Fellowship.

Author information

Authors and Affiliations

  1. School of Biological Sciences, Universiti Sains Malaysia, 11800, Gelugor, Penang, Malaysia

    Nazila Biglari & Kumar Sudesh

  2. Enteric Diseases Research Cluster, Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia (USM), 11800, Gelugor, Penang, Malaysia

    Marjan Ganjali Dashti

  3. Department of Microbiology, Masjed Soleiman Branch, Islamic Azad University, Masjed Soleiman, Iran

    Peyman Abdeshahian

  4. School of Life Science and Technology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan

    Izumi Orita & Toshiaki Fukui

Authors
  1. Nazila Biglari
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marjan Ganjali Dashti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peyman Abdeshahian
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Izumi Orita
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Toshiaki Fukui
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kumar Sudesh
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

NB certifies that she has participated full-time to make extensive contributions to conception and design, and acquisition of data, and analysis and interpretation of data for the entire content of the manuscript and writing the manuscript. MGD certifies that she has participated sufficiently in the work to take public responsibility for data analysis and writing the manuscript. PA certifies that she has participated sufficiently in the work to take public responsibility for discussion section and writing the manuscript. IO, TF give final approval of the version to be submitted and any revised version. KS certifies that he has participated in drafting the article or revising it critically for important intellectual content.

Corresponding author

Correspondence to Kumar Sudesh.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Biglari, N., Ganjali Dashti, M., Abdeshahian, P. et al. Enhancement of bioplastic polyhydroxybutyrate P(3HB) production from glucose by newly engineered strain Cupriavidus necator NSDG-GG using response surface methodology. 3 Biotech 8, 330 (2018). https://doi.org/10.1007/s13205-018-1351-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s13205-018-1351-7

Keywords

Search

Navigation