Various hydrogenases and formate-dependent hydrogen production in Citrobacter amalonaticus Y19

Abstract An isolate Citrobacter amalonaticus Y19 showed a typical mixed-acid fermentation with lactate and acetate as major end products when grown anaerobically on glucose and pyruvate, respectively. Production of hydrogen ( H 2 ) from glucose, formate, and reduced methylviologen (MV) and benzylviologen (BV) by the resting cells of Y19 indicates the presence of formate hydrogen lyase (FHL) activity and other hydrogenases. Study with subcellular fractions of Y19 exhibited that the FHL activity, dependent on soluble formate dehydrogenase activity, was detected in the broken cell extract, but not in the soluble or particulate fraction which are separated by centrifugation at 35 , 000 × g . Hydrogen production in the presence of reduced MV or BV was observed in both the soluble and particulate fractions. Uptake hydrogenase activity was observed in both the fractions when the oxidized forms of MV and BV were supplied as electron acceptor. In the soluble fraction, when formate was coupled with oxidized form of MV or BV, hydrogen production activity was recovered. These results indicate that, similar to E. coli, the strain Y19 expresses two different hydrogenases, one as the FHL complex and another as membrane-associated enzyme.

[1]  You-Kwan Oh,et al.  Fermentative biohydrogen production by a new chemoheterotrophic bacterium Citrobacter sp. Y19 , 2003 .

[2]  R. Thauer,et al.  On the Mechanism of Catalysis by a Metal‐Free Hydrogenase from Methanogenic Archaea: Enzymatic Transformation of H2 without a Metal and Its Analogy to the Chemistry of Alkanes in Superacidic Solution , 1995 .

[3]  Ji-Young Park,et al.  A new chemoheterotrophic bacterium catalyzing water-gas shift reaction , 1999, Biotechnology Letters.

[4]  Michel Frey,et al.  Crystal structure of the nickel–iron hydrogenase from Desulfovibrio gigas , 1995, Nature.

[5]  T. Kanai,et al.  Characterization of a Cytosolic NiFe-Hydrogenase from the Hyperthermophilic Archaeon Thermococcus kodakaraensis KOD1 , 2003, Journal of bacteriology.

[6]  P. Vignais,et al.  Molecular biology of microbial hydrogenases. , 2004, Current issues in molecular biology.

[7]  K. Bagramyan,et al.  The roles of hydrogenases 3 and 4, and the F0F1‐ATPase, in H2 production by Escherichia coli at alkaline and acidic pH , 2002, FEBS letters.

[8]  Maness Pc,et al.  Evidence for three distinct hydrogenase activities in Rhodospirillum rubrum. , 2001 .

[9]  R. Sawers,et al.  Differential expression of hydrogenase isoenzymes in Escherichia coli K-12: evidence for a third isoenzyme , 1985, Journal of bacteriology.

[10]  Shigeharu Tanisho,et al.  Continuous hydrogen production from molasses by fermentation using urethane foam as a support of flocks , 1995 .

[11]  D. Zannoni.  Respiration in Archaea and Bacteria , 2004, Advances in Photosynthesis and Respiration.

[12]  Lawrence Pitt,et al.  Biohydrogen production: prospects and limitations to practical application , 2004 .

[13]  J. R. Kim,et al.  Oxygen Sensitivity of Carbon Monoxide-Dependent Hydrogen Production Activity in Citrobacter sp. , 2003 .

[14]  G. Sawers,et al.  Isolation and characterization of hypophosphite‐resistant mutants of Escherichia coli: identification of the FocA protein, encoded by the pfl operon, as a putative formate transporter , 1994, Molecular microbiology.

[15]  G. Sawers.  The hydrogenases and formate dehydrogenases ofEscherichia coli , 2004, Antonie van Leeuwenhoek.

[16]  J. R. Kim,et al.  Hydrogen production by a new chemoheterotrophic bacterium Citrobacter sp. Y19 , 2002 .

[17]  B J Lemon,et al.  X-ray crystal structure of the Fe-only hydrogenase (CpI) from Clostridium pasteurianum to 1.8 angstrom resolution. , 1998, Science.

[18]  M. Inui,et al.  Enhanced Hydrogen Production from Formic Acid by Formate Hydrogen Lyase-Overexpressing Escherichia coli Strains , 2005, Applied and Environmental Microbiology.

[19]  R. Sawers,et al.  Formate and its role in hydrogen production in Escherichia coli. , 2005, Biochemical Society transactions.