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Batch, design optimization, and DNA sequencing study for continuous 1,3-propanediol production from waste glycerol by a soil-based inoculum

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Abstract

1,3-propanediol (1,3-PD) was produced with a robust fermentation process using waste glycerol feedstock from biodiesel production and a soil-based bacterial inoculum. An iterative inoculation method was developed to achieve independence from soil and selectively breed bacterial populations capable of glycerol metabolism to 1,3-PD. The inoculum showed high resistance to impurities in the feedstock. 1,3-PD selectivity and yield in batch fermentations was optimized by appropriate nutrient compositions and pH control. The batch yield of 1,3-PD was maximized to ~0.7 mol/mol for industrial glycerol which was higher than that for pure glycerin. 16S rDNA sequencing results show a systematic selective enrichment of 1,3-PD producing bacteria with iterative inoculation and subsequent process control. A statistical design of experiments was carried out on industrial glycerol batches to optimize conditions, which were used to run two continuous flow stirred-tank reactor (CSTR) experiments over a period of >500 h each. A detailed analysis of steady states at three dilution rates is presented. Enhanced specific 1,3-PD productivity was observed with faster dilution rates due to lower levels of solvent degeneration. 1,3-PD productivity, specific productivity, and yield of 1.1 g/l hr, 1.5 g/g hr, and 0.6 mol/mol of glycerol were obtained at a dilution rate of 0.1 h−1which is bettered only by pure strains in pure glycerin feeds.

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Acknowledgments

The authors gratefully acknowledge Prof. Joerg Graf and the Department of Molecular and Cell Biology, University of Connecticut, for their assistance in the 16S rDNA sequencing experiments. Portions of this work were supported by DOE Grant DE-EE0003116.

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Authors and Affiliations

  1. Institute of Materials Science, University of Connecticut, 97 N. Eagleville Rd., Storrs, CT, 06269, USA

    Baishali Kanjilal & Richard S. Parnas

  2. Department of Chemical and Biomolecular Engineering, University of Connecticut, 191 Auditorium Rd., Storrs, CT, 06269, USA

    Iman Noshadi, Eddy J. Bautista, Ranjan Srivastava & Richard S. Parnas

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  1. Baishali Kanjilal
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  2. Iman Noshadi
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  3. Eddy J. Bautista
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  4. Ranjan Srivastava
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  5. Richard S. Parnas
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Correspondence to Richard S. Parnas.

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Kanjilal, B., Noshadi, I., Bautista, E.J. et al. Batch, design optimization, and DNA sequencing study for continuous 1,3-propanediol production from waste glycerol by a soil-based inoculum. Appl Microbiol Biotechnol 99, 2105–2117 (2015). https://doi.org/10.1007/s00253-014-6259-5

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