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Isolation of microorganisms for biological detoxification of lignocellulosic hydrolysates

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Abstract

Acid pretreatment of lignocellulosic biomass releases furan and phenolic compounds, which are toxic to microorganisms used for subsequent fermentation. In this study, we isolated new microorganisms for depletion of inhibitors in lignocellulosic acid hydrolysates. A sequential enrichment strategy was used to isolate microorganisms from soil. Selection was carried out in a defined mineral medium containing a mixture of ferulic acid (5 mM), 5-hydroxymethylfurfural (5-HMF, 15 mM), and furfural (20 mM) as the carbon and energy sources, followed by an additional transfer into a corn stover hydrolysate (CSH) prepared using dilute acid. Subsequently, based on stable growth on these substrates, six isolates—including five bacteria related to Methylobacterium extorquens, Pseudomonas sp, Flavobacterium indologenes, Acinetobacter sp., Arthrobacter aurescens, and one fungus, Coniochaeta ligniaria—were chosen. All six isolates depleted toxic compounds from defined medium, but only C. ligniaria C8 (NRRL 30616) was effective at eliminating furfural and 5-HMF from CSH. C. ligniaria NRRL 30616 may be useful in developing a bioprocess for inhibitor abatement in the conversion of lignocellulosic biomass to fuels and chemicals.

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Acknowledgements

We wish to thank Dr. Clete Kurtzman and Dr. Donald Wicklow for help identifying fungal strain C8, and Dr. Terrence Whitehead for providing primers for amplification of bacterial rDNA. We thank Ms. Patricia O'Bryan and Mr. Loren Iten for excellent technical assistance, and Dr. Evi Weber for helpful discussion. This work was supported by ARS-USDA through cooperative research agreement 3620-41000-084-03

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Author notes

  1. R. J. Bothast

    Present address: National Corn-to-Ethanol Research Pilot Plant, Southern Illinois University—Edwardsville, Edwardsville, IL 62025, USA

Authors and Affiliations

  1. Área de Microbiología, Departamento de Biología Aplicada, CITE II-B, Universidad de Almería, La Cañada de San Urbano, 04120, Almeria, Spain

    M. J. López & J. Moreno

  2. Fermentation Biotechnology Research Unit, National Center for Agricultural Utilization Research, Agricultural Research Service, US Department of Agriculture, 1815N University Street, Peoria, IL 61604, USA

    N. N. Nichols, B. S. Dien & R. J. Bothast

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  1. M. J. López
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Correspondence to M. J. López.

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Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable

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López, M.J., Nichols, N.N., Dien, B.S. et al. Isolation of microorganisms for biological detoxification of lignocellulosic hydrolysates. Appl Microbiol Biotechnol 64, 125–131 (2004). https://doi.org/10.1007/s00253-003-1401-9

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  • DOI: https://doi.org/10.1007/s00253-003-1401-9

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