3 Biotech

, 9:41 | Cite as

Mixed fermentation of Aspergillus niger and Candida shehatae to produce bioethanol with ionic-liquid-pretreated bagasse

  • Zaiqiang WuEmail author
Original Article


In this study, bagasse was pretreated with ionic liquid (IL) 1-butyl-3-methylimidazolium chloride ([Bmim]Cl) and 1% NaOH solution for initial activation of bagasse. A mixed fermentation of treated bagasse by Aspergillus niger and Candida shehatae showed the optimal conditions with the addition of C. shehatae 12 h later at a 1:1 proportion to A. niger. To further improve the ethanol production and obtain optimal fermentation conditions, a Plackett–Burman design was applied to screen the significant formulation and process variables. The optimal ethanol fermentation conditions with IL pretreated bagasse were determined using response surface methodology by Box–Behnken design. Three variables “initial pH, (NH4)2SO4, fermentation time” were regarded as significant factors in the optimization study. The resulting optimum fermentation conditions for bioethanol was identified as: initial pH of 5.89, (NH4)2SO4 concentration of 0.40 g/50 mL, and fermentation time of 3.60 days. The verification experimental ethanol concentration was 8.14 g/L, which agreed with the predicted value. An enhancement of approximately 153.58% compared with initial fermentation conditions in ethanol production was found using optimized conditions. It demonstrated that optimization methodology had a positive effect on the improvement of ethanol production. Under the optimal fermentation medium and conditions, the ethanol production with IL-pretreated bagasse and untreated bagasse was 8.14 g/L and 5.03 g/L, respectively, which exhibited 62% increase, compared to initial conditions with production of 3.21 g/L and 2.67 g/L, respectively, which displayed 20% increase. Both under optimal and original fermentation conditions, compared to the fermentation medium with untreated bagasse, all the results indicated that IL-pretreated bagasse resulted in higher ethanol production than untreated bagasse, demonstrating that IL-pretreated bagasse successfully increased the ethanol production in the mixed fermentation by A. niger and C. shehatae.


Ionic liquid Mixed fermentation Plackett–Burman design Box–Behnken design Response surface methodology 



The author is grateful for the support of all the members from Molecular Metabolic center of Nanjing University of Science and Technology.

Compliance with ethical standards

Conflict of interest

We confirm that this manuscript has neither been published elsewhere nor was under consideration by another journal. All authors approved this manuscript and agree with its submission to your journal. The authors have no conflicts of interest.

Supplementary material

13205_2019_1570_MOESM1_ESM.docx (15 kb)
Supplementary material 1 (DOCX 15 KB)


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© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.Center for Molecular Metabolism, School of Environmental and Biological EngineeringNanjing University of Science and TechnologyNanjingChina

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