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Molecular analysis of the anaerobic succinate degradation pathway in Clostridium kluyveri
A region of genomic DNA from Clostridium kluyveri was cloned in Escherichia coli by a screening strategy which was based on heterologous expression of the clostridial 4-hydroxybutyrate dehydrogenase gene, and similarities to the adhE (aad) gene products from E. coli were revealed.
Purification and characterization of a coenzyme-A-dependent succinate-semialdehyde dehydrogenase from Clostridium kluyveri.
Cell extracts of Clostridium kluyveri, grown on ethanol plus succinate contained a succinyl-CoA:CoA transferase, a coenzyme-A-dependent succinate-semialdehyde dehydrogenase, and a NAD(+)-dependent 4-hydroxybutyrate dehydrogenases, which suggest a ping-pong mechanism.
Various functions of selenols and thiols in anaerobic Gram‐positive, amino acids‐utilizing bacteria
Electron transfer reactions for the reduction of glycine in Eubacterium acidaminophilum involve many selenocysteine (U)‐ and thiol‐containing proteins, as shown by biochemical and molecular analysis.
A novel fusion protein system for the production of native human pepsinogen in the bacterial periplasm.
Human pepsinogen was fused to E. coli ecotin (E. coli trypsin inhibitor), which is a periplasmic homodimeric protein of 142 amino acids per monomer containing one disulfide bridge, and was expressed in pTrc99a.
Use of enhanced green fluorescent protein to determine pepsin at high sensitivity.
A fluorometric assay for pepsin and pepinogen was developed using enhanced green fluorescent protein (EGFP) as a substrate, which revealed distinct cleavage sites, as analyzed by SDS-PAGE.
Biosynthesis of poly(4-hydroxybutyric acid) by recombinant strains of Escherichia coli.
The homopolyester poly(4HB) was also accumulated during cultivation of these strains in M9 mineral salts medium containing glucose plus 4-hydroxybutyric acid as carbon sources and if the cultures were not sufficiently supplied with oxygen.
A selDABC cluster for selenocysteine incorporation in Eubacterium acidaminophilum
As the benzyl viologen-dependent formate dehydrogenase activity in these strains after anaerobic growth in the presence of formate is determined, Transformation of this E. coli deletion mutant with selB alone was not sufficient to restore formate dehydration activity.
Factors and Selenocysteine Insertion Sequence Requirements for the Synthesis of Selenoproteins from a Gram-Positive Anaerobe in Escherichia coli
UGA readthrough of SECIS elements present in Desulfomicrobium baculatum (hydV), Treponema denticola (selD), and Campylobacter jejuni (selW-like gene) was considerably enhanced in the presence of E. acidaminophilum selB and selC, indicating recognition of theseSECIS elements and might open new perspectives for heterologous selenoprotein synthesis in E. coli.
Succinate-ethanol fermentation in Clostridium kluyveri: purification and characterisation of 4-hydroxybutyryl-CoA dehydratase/vinylacetyl-CoA Δ3-Δ2-isomerase
Anaerobically prepared cell extracts of Clostridium kluyveri grown on succinate plus ethanol contained high amounts of 4-hydroxybutyryl-CoA dehydratase, which catalyzes the reversible dehydration of
Substrate-specific selenoprotein B of glycine reductase from Eubacterium acidaminophilum. Biochemical and molecular analysis.
The substrate-specific selenoprotein B of glycine reductase (PBglycine) from Eubacterium acidaminophilum was purified and characterized, and peptide sequence analyses suggest that grdE encodes a proprotein which is cleaved into the previously sequenced N-terminal 25-kDa (beta) subunit and a 22-k da (alpha) sub unit of PBglycines.