Identification of the electron transfer flavoprotein as an upregulated enzyme in the benzoate utilization of Desulfotignum balticum.
Desulfotignum balticum utilizes benzoate coupled to sulfate reduction. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis was conducted to detect proteins that increased more after growth on benzoate than on butyrate. A comparison of proteins on 2D gels showed that at least six proteins were expressed. The N-terminal sequences of three proteins exhibited significant identities with the alpha and beta subunits of electron transfer flavoprotein (ETF) from anaerobic aromatic-degraders. By sequence analysis of the fosmid clone insert (37,590 bp) containing the genes encoding the ETF subunits, we identified three genes, whose deduced amino acid sequences showed 58%, 74%, and 62% identity with those of Gmet_2267 (Fe-S oxidoreductase), Gmet_2266 (ETF beta subunit), and Gmet_2265 (ETF alpha subunit) respectively, which exist within the 300-kb genomic island of aromatic-degradation genes from Geobacter metallireducens GS-15. The genes encoding ETF subunits found in this study were upregulated in benzoate utilization.
Habe
Hiroshi
H
Research Institute for Innovations in Sustainable Chemistry, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan. hiroshi.habe@aist.go.jp
Kobuna
Akinori
A
Hosoda
Akifumi
A
Kosaka
Tomoyuki
T
Endoh
Takayuki
T
Tamura
Hiroto
H
Yamane
Hisakazu
H
Nojiri
Hideaki
H
Omori
Toshio
T
Watanabe
Kazuya
K
D000595:Amino Acid Sequence; D001565:Benzoates / Q000494:pharmacology*; D002244:Carbon / Q000737:chemistry; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D020564:Deltaproteobacteria / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D044927:Electron-Transferring Flavoproteins / Q000235:genetics* / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D015723:Gene Library; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D015854:Up-Regulation / Q000187:drug effects*
D001565:Benzoates; D004269:DNA, Bacterial; D044927:Electron-Transferring Flavoproteins; D010455:Peptides; D021122:Protein Subunits; D002244:Carbon