10.1021/es1002023.s001 Amy V. Callaghan Amy V. Callaghan Irene A. Davidova Irene A. Davidova Kristen Savage-Ashlock Kristen Savage-Ashlock Victoria A. Parisi Victoria A. Parisi Lisa M. Gieg Lisa M. Gieg Joseph M. Suflita Joseph M. Suflita Jerome J. Kukor Jerome J. Kukor Boris Wawrik Boris Wawrik Diversity of Benzyl- and Alkylsuccinate Synthase Genes in Hydrocarbon-Impacted Environments and Enrichment Cultures American Chemical Society 2010 alkane degradation ALDC aliphatic hydrocarbon biodegradation PCR Alkylsuccinate Synthase Genes ASS assA genes BSS DNA biomarker 2010-10-01 00:00:00 Journal contribution https://acs.figshare.com/articles/journal_contribution/Diversity_of_Benzyl_and_Alkylsuccinate_Synthase_Genes_in_Hydrocarbon_Impacted_Environments_and_Enrichment_Cultures/2725210 Hydrocarbon-degrading microorganisms play an important role in the natural attenuation of spilled petroleum in a variety of anoxic environments. The role of benzylsuccinate synthase (BSS) in aromatic hydrocarbon degradation and its use as a biomarker for field investigations are well documented. The recent discovery of alkylsuccinate synthase (ASS) allows the opportunity to test whether its encoding gene, <i>assA</i>, can serve as a comparable biomarker of anaerobic alkane degradation. Degenerate <i>assA</i>- and <i>bssA</i>-targeted PCR primers were designed in order to survey the diversity of genes associated with aromatic and aliphatic hydrocarbon biodegradation in petroleum-impacted environments and enrichment cultures. DNA was extracted from an anaerobic alkane-degrading isolate (<i>Desulfoglaeba alkenexedens</i> ALDC), hydrocarbon-contaminated river and aquifer sediments, a paraffin-degrading enrichment, and a propane-utilizing mixed culture. Partial <i>assA</i> and <i>bssA</i> genes were PCR amplified, cloned, and sequenced, yielding several novel clades of <i>assA</i> genes. These data expand the range of alkane-degrading conditions for which relevant gene sequences are available and indicate that considerable diversity of <i>assA</i> genes can be found in hydrocarbon-impacted environments. The detection of genes associated with anaerobic alkane degradation in conjunction with the in situ detection of alkylsuccinate metabolites was also demonstrated. Comparable molecular signals of <i>assA</i>/<i>bssA</i> were not found when environmental metagenome databases of uncontaminated sites were searched. These data confirm that the <i>assA</i> gene is a useful biomarker for anaerobic alkane metabolism.