Open Access Research

Biological conversion assay using Clostridium phytofermentans to estimate plant feedstock quality

Scott J Lee12, Thomas A Warnick3, Sivakumar Pattathil5, Jesús G Alvelo-Maurosa3, Michelle J Serapiglia6, Heather McCormick5, Virginia Brown5, Naomi F Young1, Danny J Schnell4, Lawrence B Smart6, Michael G Hahn5, Jeffrey F Pedersen7, Susan B Leschine3 and Samuel P Hazen1*

Author Affiliations

1 Biology Department, University of Massachusetts, Amherst, MA, USA

2 Plant Biology Graduate Program, University of Massachusetts, Amherst, MA, USA

3 Department of Microbiology, University of Massachusetts, Amherst, MA, USA

4 Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA, USA

5 BioEnergy Science Center, Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA

6 Department of Horticulture, Cornell University, Geneva, NY, USA

7 USDA-ARS, Grain, Forage, and Bioenergy Research, University of Nebraska-Lincoln, Lincoln, NE, USA

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Biotechnology for Biofuels 2012, 5:5  doi:10.1186/1754-6834-5-5

Published: 8 February 2012



There is currently considerable interest in developing renewable sources of energy. One strategy is the biological conversion of plant biomass to liquid transportation fuel. Several technical hurdles impinge upon the economic feasibility of this strategy, including the development of energy crops amenable to facile deconstruction. Reliable assays to characterize feedstock quality are needed to measure the effects of pre-treatment and processing and of the plant and microbial genetic diversity that influence bioconversion efficiency.


We used the anaerobic bacterium Clostridium phytofermentans to develop a robust assay for biomass digestibility and conversion to biofuels. The assay utilizes the ability of the microbe to convert biomass directly into ethanol with little or no pre-treatment. Plant samples were added to an anaerobic minimal medium and inoculated with C. phytofermentans, incubated for 3 days, after which the culture supernatant was analyzed for ethanol concentration. The assay detected significant differences in the supernatant ethanol from wild-type sorghum compared with brown midrib sorghum mutants previously shown to be highly digestible. Compositional analysis of the biomass before and after inoculation suggested that differences in xylan metabolism were partly responsible for the differences in ethanol yields. Additionally, we characterized the natural genetic variation for conversion efficiency in Brachypodium distachyon and shrub willow (Salix spp.).


Our results agree with those from previous studies of lignin mutants using enzymatic saccharification-based approaches. However, the use of C. phytofermentans takes into consideration specific organismal interactions, which will be crucial for simultaneous saccharification fermentation or consolidated bioprocessing. The ability to detect such phenotypic variation facilitates the genetic analysis of mechanisms underlying plant feedstock quality.