Evaluation of Cellulose as a Substrate for Hydrocarbon Fuel Production by Ascocoryne sarcoides (NRRL 50072)


The fungal endophyte, Ascocoryne sarcoides, produced aviation, gasoline and diesel-relevant hydrocarbons when grown on multiple substrates including cellulose as the sole carbon source. Substrate, growth stage, culturing pH, temperature and medium composition were statistically significant factors for the type and quantity of hydrocarbons produced. Gasoline range (C5-C12), aviation range (C8-C16) and diesel range (C9-C36) organics were detected in all cultured media. Numerous non-oxygenated hydrocarbons were produced such as isopentane, 3,3-dimethyl hexane and d-limonene during exponential growth phase. Growth on cellulose at 23°C and pH 5.8 produced the highest overall yield of fuel range organics (105 mg * g·biomass-1). A change in metabolism was seen in late stationary phase from catabolism of cellulose to potential oxidation of hydrocarbons resulting in the production of more oxygenated compounds with longer carbon chain length and fewer fuel-related compounds. The results outline rational strategies for controlling the composition of the fuel-like compounds by changing culturing parameters.

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Mallette, N. , M. Pankratz, E. , E. Parker, A. , A. Strobel, G. , C. Busse, S. , P. Carlson, R. and M. Peyton, B. (2014) Evaluation of Cellulose as a Substrate for Hydrocarbon Fuel Production by Ascocoryne sarcoides (NRRL 50072). Journal of Sustainable Bioenergy Systems, 4, 33-49. doi: 10.4236/jsbs.2014.41004.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Carroll, A. and Somerville, C. (2009) Cellulosic Biofuels. Annual Review of Plant Biology, 60, 165-182.
[2] Perlack, R.D., Wright, L.L., Turhollow, A.F., Graham, R.L., Stokes, B.J. and Erbach, D.C. (2005) Biomass as Feedstock for a Bioenergy and Bioproducts Industry: The Technical Feasibility of a Billion-Ton Annual Feedstock Supply. Oak Ridge National Laboratory /USDOE/USDA, Oak Ridge. http://dx.doi.org/10.2172/885984
[3] Dien, B.S., Cotta, M.A. and Jeffries, T.W. (2003) Bacteria Engineered for Fuel Ethanol Production: Current Status. Applied Microbiology and Biotechnology, 63, 258-266. http://dx.doi.org/10.1007/s00253-003-1444-y
[4] Ragauskas, A.J., Williams, C.K., Davison, B.H., Britovsek, G., Cairney, J., Eckert, C.A., et al. (2006) The Path Forward for Biofuels and Biomaterials. Science, 311, 484-489. http://dx.doi.org/10.1126/science.1114736
[5] Strobel, G.A., Knighton, B., Kluck, K., Ren, Y., Livinghouse, T., Griffin, M., et al. (2008) The Production of MycoDiesel Hydrocarbons and Their Derivatives by the Endophytic Fungus Gliocladium roseum (NRRL 50072). Microbiology, 154, 3319-3328. http://dx.doi.org/10.1099/mic.0.2008/022186-0
[6] Griffin, M.A., Spakowicz, D.J., Gianoulis, T.A. and Strobel, S.A. (2010) Volatile Organic Compound Production by Organisms in the Genus Ascocoryne and a Re-Evaluation of Myco-Diesel Production by NRRL 50072. Microbiology, 156, 3814-3829. http://dx.doi.org/10.1099/mic.0.041327-0
[7] Mallette, N.D., Knighton, W.B., Strobel, G.A., Carlson, R.P., Peyton, B.M. (2012) Resolution of Volatile Fuel Compound Profiles from Ascocoryne Sarcoides: A Comparison by Proton Transfer Reaction-Mass Spectrometry and Solid Phase Microextraction Gas Chromatography-Mass Spectrometry. AMB Express, 2, 23.
[8] Mends, M.T., Yu, E., Strobel, G.A., Riyaz-Ul-Hassan, S., Booth, E., Geary, B., et al. (2012) An Endophytic Nodulisporium sp. Producing Volatile Organic Compounds Having Bioactivity and Fuel Potential. Journal of Petroleum & Environmental Biotechnology, 3. http://dx.doi.org/10.4172/2157-7463.1000117
[9] Morath, S.U., Hung, R. and Bennett, J.W. (2012) Fungal Volatile Organic Compounds: A Review with Emphasis on Their Biotechnological Potential. Fungal Biology Reviews, 26, 73-83. http://dx.doi.org/10.1016/j.fbr.2012.07.001
[10] Yuan, Z.-L., Chen, Y.-C., Xu, B.-G. and Zhang, C.-L. (2012) Current Perspectives on the Volatile-Producing Fungal Endophytes. Critical Reviews in Biotechnology, 32, 363-373. http://dx.doi.org/10.3109/07388551.2011.651429
[11] Gianoulis, T.A., Griffin, M.A., Spakowicz, D.J., Dunican, B.F., Alpha, C.J., Sboner, A., et al. (2012) Genomic Analysis of the Hydrocarbon-Producing, Cellulolytic, Endophytic Fungus Ascocoryne sarcoides. PLOS Genetics, 8, Article ID: e1002558. http://dx.doi.org/10.1371/journal.pgen.1002558
[12] Stewart, J.C. and Parry, J.B. (1981) Factors Influencing the Production of Cellulase by Aspergillus fumigatus (Fresenius). Journal of General Microbiology, 125, 33-39. http://dx.doi.org/10.1099/00221287-125-1-33
[13] Dagbagli, S. and Goksungur, Y. (2008) Optimization of Beta-Galactosidase Production Using Kluyveromyces lactis NRRL Y-8279 by Response Surface Methodology. Electronic Journal of Biotechnology, 11.
[14] Bates, D., Maechler, M. and Bolker, B. (2013) lme4: Linear Mixed-Effects Models Using S4 Classes. R Package Version 0.999999-2.
[15] R Core Team (2013) R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna.
[16] Booth, E., Strobel, G., Knighton, B., Sears, J., Geary, B. and Avci, R. (2011) A Rapid Column Technique for Trapping and Collecting of Volatile Fungal Hydrocarbons and Hydrocarbon Derivatives. Biotechnology Letters, 33, 1963-1972.
[17] Linstrom, P.J. and Mallard, W.G. (2011) NIST Chemistry WebBook. NIST Standard Reference Database Number 69, National Institute of Standards and Technology, Gaithersburg.
[18] Stinson, M., Ezra, D., Hess, W.M., Sears, J. and Strobel, G. (2003) An endophytic Gliocladium sp. of Eucryphia Cordifolia Producing Selective Volatile Antimicrobial Compounds. Plant Science, 165, 913-922.
[19] Ul-Hassan, S.R., Strobel, G.A., Booth, E., Knighton, B., Floerchinger, C. and Sears, J. (2012) Modulation of Volatile Organic Compound Formation in the Mycodiesel-Producing Endophyte Hypoxylon sp. CI-4. Microbiology, 158, 465473. http://dx.doi.org/10.1099/mic.0.054643-0
[20] Speight, J.G. (2002) Handbook of Petroleum Product Analysis—Knovel. John Wiley & Sons, Inc., Hoboken.
[21] Yoon, S.H. and Lee, C.S. (2012) Effect of Undiluted Bioethanol on Combustion and Emissions Reduction in a Si Engine at Various Charge Air Conditions. Fuel, 97, 887-890. http://dx.doi.org/10.1016/j.fuel.2012.02.001
[22] Blakey, S., Rye, L. and Wilson, C.W. (2011) Aviation Gas Turbine Alternative Fuels: A Review. Proceedings of the Combustion Institute, Proceedings of the Combustion Institute, 33, 2863-2885.
[23] Ciapetta, F.G. and Wallace, D.N. (1972) Catalytic Naphtha Reforming. Catalysis Reviews, 5, 67-158.
[24] Ghosh, P., Hickey, K.J. and Jaffe, S.B. (2005) Development of a Detailed Gasoline Composition-Based Octane Model. Industrial & Engineering Chemistry Research, 45, 337-345. http://dx.doi.org/10.1021/ie050811h

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