, Volume 22, Issue 2, pp 309–320 | Cite as

Effect of nitrogen source on methanol oxidation and genetic diversity of methylotrophic mixed cultures enriched from pulp and paper mill biofilms

Original Paper


Methanol-oxidizing bacteria may play an important role in the development and use of biological treatment systems for the removal of methanol from industrial effluents. Optimization of methanol degradation potential in such systems is contingent on availability of nutrients, such as nitrogen, in the most favorable form and concentration. To that end, this study examined the variation in growth, methanol degradation, and bacterial diversity of two mixed methylotrophic cultures that were provided nitrogen either as ammonium or nitrate and in three different concentrations. Methanol-degrading cultures were enriched from biofilms sampled at a pulp and paper mill and grown in liquid batch culture with methanol as the only carbon source and either ammonium or nitrate as the only added nitrogen source. Results indicate that growth and methanol removal of the mixed cultures increase directly with increased nitrogen, added in either form. However, methanol removal and bacterial diversity, as observed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR–DGGE) methods, were higher when using nitrate as the nitrogen source for enrichment and growth, rather than ammonium. Based on results described here, nitrate may potentially be a better nitrogen source when enriching or working with mixed methylotrophic cultures, and possibly more effective when used as a nutrient addition to biofilters.


Methylotroph Nitrogen source Methanol biofilter Bacterial diversity 



This study was supported by the Department of Energy, Award Number: DE-FC36-03ID14437. The authors acknowledge and gratefully thank Adriana Pacheco, Rebecca McLarty, Shweta Patole, Michael Friedlander, Mauricio Arias, Greg Babbitt, and Jennifer Stokke (Unviersity of Florida) for assistance in laboratory data collection and sample analysis; Ashok Jain, Jim Stainfield, and Karen Mentz (NCASI) for technical advisement and data analysis; and Timothy McKelvey, Chet Thompson, Cecile Hance, and Myra Carpenter (industry representatives) for assistance in collection of biofilm and other process samples and process information.


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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Golisano Institute for SustainabilityRochester Institute of TechnologyRochesterUSA
  2. 2.Department of Environmental Engineering SciencesUniversity of FloridaGainesvilleUSA

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