New FeFe-hydrogenase genes identified in a metagenomic fosmid library from a municipal wastewater treatment plant as revealed by high-throughput sequencing

Autor(es)

Tomazetto, Geizecler ^[1] ; Wibberg, Daniel ^[2] ; Schlueter, Andreas ^[2] ; Oliveira, Valeria M. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Campinas, Res Ctr Chem Biol & Agr CPQBA, Div Microbial Resources, BR-13081970 Campinas, SP - Brazil

^[2] Univ Bielefeld, CeBiTec, Inst Genome Res & Syst Biol, D-33615 Bielefeld - Germany

• Tipo de documento: Artigo Científico
• Fonte: Research in Microbiology; v. 166, n. 1, p. 9-19, JAN 2015.
• Citações Web of Science: 3

Resumo

A fosmid metagenomic library was constructed with total community DNA obtained from a municipal wastewater treatment plant (MWWTP), with the aim of identifying new FeFe-hydrogenase genes encoding the enzymes most important for hydrogen metabolism. The dataset generated by pyrosequencing of a fosmid library was mined to identify environmental gene tags (EGTs) assigned to FeFe-hydrogenase. The majority of EGTs representing FeFe-hydrogenase genes were affiliated with the class Clostridia, suggesting that this group is the main hydrogen producer in the MWWTP analyzed. Based on assembled sequences, three FeFe-hydrogenase genes were predicted based on detection of the L2 motif (MPCxxKicxE) in the encoded gene product, confirming true FeFe-hydrogenase sequences. These sequences were used to design specific primers to detect fosmids encoding FeFe-hydrogenase genes predicted from the dataset. Three identified fosmids were completely sequenced. The cloned genomic fragments within these fosmids are closely related to members of the Spirochaetaceae, Bacteroidales and Firmicutes, and their FeFe-hydrogenase sequences are characterized by the structure type M3, which is common to clostridial enzymes. FeFe-hydrogenase sequences found in this study represent hitherto undetected sequences, indicating the high genetic diversity regarding these enzymes in MWWTP. Results suggest that MWWTP have to be considered as reservoirs for new FeFe-hydrogenase genes. (C) 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved. (AU)

• Processo FAPESP: 09/01823-5 - Diversidade e prospecção de metagenoma microbiano em fermentadores de biogás produzindo H2
• Beneficiário: Geizecler Tomazetto
• Modalidade de apoio: Bolsas no Brasil - Doutorado