Environmental Science and Pollution Research

, Volume 24, Issue 9, pp 8790–8804 | Cite as

Biohydrogen production from sugarcane bagasse hydrolysate: effects of pH, S/X, Fe2+, and magnetite nanoparticles

  • Karen Reddy
  • Mahmoud Nasr
  • Sheena Kumari
  • Santhosh Kumar
  • Sanjay Kumar Gupta
  • Abimbola Motunrayo Enitan
  • Faizal Bux
Research Article


Batch dark fermentation experiments were conducted to investigate the effects of initial pH, substrate-to-biomass (S/X) ratio, and concentrations of Fe2+ and magnetite nanoparticles on biohydrogen production from sugarcane bagasse (SCB) hydrolysate. By applying the response surface methodology, the optimum condition of steam-acid hydrolysis was 0.64% (v/v) H2SO4 for 55.7 min, which obtained a sugar yield of 274 mg g−1. The maximum hydrogen yield (HY) of 0.874 mol (mol glucose−1) was detected at the optimum pH of 5.0 and S/X ratio of 0.5 g chemical oxygen demand (COD, g VSS−1). The addition of Fe2+ 200 mg L−1 and magnetite nanoparticles 200 mg L−1 to the inoculum enhanced the HY by 62.1% and 69.6%, respectively. The kinetics of hydrogen production was estimated by fitting the experimental data to the modified Gompertz model. The inhibitory effects of adding Fe2+ and magnetite nanoparticles to the fermentative hydrogen production were examined by applying Andrew’s inhibition model. COD mass balance and full stoichiometric reactions, including soluble metabolic products, cell synthesis, and H2 production, indicated the reliability of the experimental results. A qPCR-based analysis was conducted to assess the microbial community structure using Enterobacteriaceae, Clostridium spp., and hydrogenase-specific gene activity. Results from the microbial analysis revealed the dominance of hydrogen producers in the inoculum immobilized on magnetite nanoparticles, followed by the inoculum supplemented with Fe2+ concentration.

Graphical abstract


Biohydrogen Dark fermentation Full stoichiometry Hydrogen-producing microbes Magnetite nanoparticles Sugarcane bagasse hydrolysate 



The authors would like to acknowledge the National Research Foundation of South Africa and the Durban University of Technology for providing financial assistance.

Supplementary material

11356_2017_8560_MOESM1_ESM.docx (3.1 mb)
ESM 1 (DOCX 3163 kb)
11356_2017_8560_MOESM2_ESM.docx (23 kb)
ESM 2 (DOCX 22 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Karen Reddy
    • 1
  • Mahmoud Nasr
    • 2
  • Sheena Kumari
    • 1
  • Santhosh Kumar
    • 3
  • Sanjay Kumar Gupta
    • 1
  • Abimbola Motunrayo Enitan
    • 1
  • Faizal Bux
    • 1
  1. 1.Institute for Water and Wastewater TechnologyDurban University of TechnologyDurbanSouth Africa
  2. 2.Sanitary Engineering Department, Faculty of EngineeringAlexandria UniversityAlexandriaEgypt
  3. 3.Department of Biotechnology and Food TechnologyDurban University of TechnologyDurbanSouth Africa

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