Open AccessThe F 420 H 2 ‐dehydrogenase from Methanolobus tindarius : cloning of the ffd operon and expression of the genes in Escherichia coli
Author(s) -
Westenberg David J,
Braune Annett,
Ruppert Claudia,
Müller Volker,
Herzberg Christina,
Gottschalk Gerhard,
Blaut Michael
Publication year - 1999
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1999.tb13399.x
Subject(s) - operon , escherichia coli , microbiology and biotechnology , gene , biology , open reading frame , nadh dehydrogenase , nucleic acid sequence , primer extension , biochemistry , protein subunit , peptide sequence , messenger rna
The membrane‐bound F 420 H 2 ‐dehydrogenase from the methylotrophic methanogen Methanolobus tindarius oxidizes reduced coenzyme F 420 and feeds the electrons into an energy‐conserving electron transport chain. Based on the N‐terminal amino acid sequence of the 40‐kDa subunit of F 420 H 2 ‐dehydrogenase the corresponding gene ffdB was detected in chromosomal DNA of M. tindarius . Sequence analysis, primer extension, and RT‐PCR experiments indicated that ffdB is part of an operon harboring three additional open reading frames ( ffdA, ffdC, ffdD ). The corresponding mRNA transcript and transcription start sites were determined. All four genes could be heterologously expressed in Escherichia coli .