Expression of different types of [FeFe]-hydrogenase genes in bacteria isolated from a population of a bio-hydrogen pilot-scale plant

Morra, Simone; Arizzi, Mariaconcetta; Allegra, Paola; Licata, B. La; Sagnelli, F.; Zitella, P.; Gilardi, Gianfranco; Valetti, Francesca

doi:10.1016/j.ijhydene.2014.04.009

Cited by 45 publications

(47 citation statements)

References 70 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…the exploitation of a novel hydrogenase recently isolated from a highly productive hydrogen pilot-plant fed with waste material [22,23], is strengthening the efforts towards a systematic identification of ideal biocatalysts to grant sustainable energy production. 35 …”

Section: Discussionmentioning

confidence: 99%

“…Recombinant expression of the hydrogenase CpHydA from Clostridium perfringens SM09 [22] was performed in E. coli 30 Rosetta2(DE3) as previously described [31] upon transformation of the competent cells with plasmid pECPF2655, obtained by ligating the entire coding sequence of CPF_2655 into the empty expression vector pECr1 [23,24] between NdeI and XhoI sites. After the induction, cells were incubated over night under pure argon flow to maintain anaerobic conditions in a water bath at 30°C.…”

Section: Recombinant Expression and Purificationmentioning

confidence: 99%

“…Conversely, stationary electrodes are easier to use and can be exploited for working devices. In this work, a novel [FeFe]-hydrogenase CpHydA (encoded by CPF_2655 gene) from a Clostridium perfringens strain newly isolated in a pilot plant with high hydrogen productivity [21][22][23] was immobilized by adsorption on stationary TiO2 electrodes. The heterologous expression in E. coli was achieved thanks to a previously established system [23,24] and it allowed to obtain a pure and active protein in good yields.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Hydrogen production at high Faradaic efficiency by a bio-electrode based on TiO2 adsorption of a new [FeFe]-hydrogenase from Clostridium perfringens

Morra

Valetti

Sarasso

et al. 2015

Bioelectrochemistry

Self Cite

View full text Add to dashboard Cite

The [FeFe]-hydrogenase CpHydA from Clostridium perfringens was immobilised by adsorption on anatase TiO2 electrodes for clean hydrogen production. The immobilized enzyme proved to perform direct electron transfer to and from the electrode surface and catalyses both H2 oxidation (H2 uptake) and H2 production (H2 evolution) with a current density for H2 20 evolution of about 2 mA cm -1 . The TiO2/CpHydA bioelectrode remained active for several days upon storage and when a reducing potential was set, H2 evolution occurred with a mean Faradaic efficiency of 98%. The high turnover frequency of H2 production and the tight coupling of electron transfer, resulting in a Faradaic efficiency close to 100%, support the exploitation of the novel TiO2/CpHydA stationary electrode as a powerful device for H2 25 production.2

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Recombinant Expression and Purificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Hydrogen production at high Faradaic efficiency by a bio-electrode based on TiO2 adsorption of a new [FeFe]-hydrogenase from Clostridium perfringens

Morra

Valetti

Sarasso

et al. 2015

Bioelectrochemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…In bioprocess, quantitative monitoring of hydrogenase enzymes is important and can provide more in-sight on the process performance. Several groups have described tools for hydrogenase monitoring at DNA and RNA levels and reported how quantitative analyses of hydrogenases can reveal important clues about the bioprocess performance8910. Previously our group reported the means to quantify Clostridium butyricum [Fe-Fe] hydrogenase at gene and transcript levels in an open bioprocess system10.…”

mentioning

confidence: 99%

“…The relationship between H 2 production kinetics and gene transcript levels of several Clostridium spp. isolated from continuous stirred tank reactor was investigated by Morra et al 9. The authors reported similar transcriptional profiles, but different H 2 evolution kinetics among the isolates indicating a post-transcriptional down regulation of clostridial hydrogenases9.…”

mentioning

confidence: 99%

Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases

Mangayil

Karp

Lamminmäki

et al. 2016

Sci Rep

View full text Add to dashboard Cite

Biological hydrogen production is based on activity of specific enzymes called hydrogenases. Hydrogenases are oxygen sensitive metalloenzymes containing Ni and/or Fe atoms at the active site, catalyzing reversible reduction of protons. Generally, [Fe-Fe] hydrogenases prefer proton reduction to molecular hydrogen, a potential energy carrier molecule that can be produced by bioprocesses in sustainable manner. Thus, monitoring tools have been developed to study the relationship between [Fe-Fe] hydrogenases and biohydrogen production in bioreactors at DNA and RNA levels. In the present study, novel molecular tools are introduced for quantitative monitoring of clostridial [Fe-Fe] hydrogenases at the protein level. Aerobic and anaerobic biopanning (for inactive and active [Fe-Fe] hydrogenase, respectively) of phage displayed single-chain variable fragment (scFv) antibody libraries aided in isolating nine potential scFvs. The enriched antibodies demonstrated high specificity towards Clostridium spp. [Fe-Fe] hydrogenases allowing detection from pure and mixed cultures. Additionally, the antibodies showed different binding characteristics towards hydrogenase catalytic states, providing a possible means for functional detection of clostridial [Fe-Fe] hydrogenases. From hydrogenase-antibody interaction studies we observed that though antibody binding reduced the enzyme catalytic activity, it facilitated to retain hydrogen evolution from oxygen exposed hydrogenases.

show abstract

Isolation and characterization of a new [FeFe]‐hydrogenase from Clostridium perfringens

Morra

Mongili

Maurelli

et al. 2015

Biotech and App Biochem

Self Cite

View full text Add to dashboard Cite

This paper reports the first characterization of an [FeFe]-hydrogenase from a Clostridium perfringens strain previously isolated in our laboratory from a pilot-scale bio-hydrogen plant that efficiently produces H2 from waste biomasses. On the basis of sequence analysis, the enzyme is a monomer formed by four domains hosting various iron-sulfur centres involved in electron transfer and the catalytic center H-cluster. After recombinant expression in Escherichia coli, the purified protein catalyzes H2 evolution at high rate of 1645 ± 16 s(-1) . The optimal conditions for catalysis are in the pH range 6.5-8.0 and at the temperature of 50 °C. EPR spectroscopy showed that the H-cluster of the oxidized enzyme displays a spectrum coherent with the Hox state, whereas the CO-inhibited enzyme has a spectrum coherent with the Hox -CO state. FTIR spectroscopy showed that the purified enzyme is composed of a mixture of redox states, with a prevalence of the Hox ; upon reduction with H2 , vibrational modes assigned to the Hred state were more abundant, whereas binding of exogenous CO resulted in a spectrum assigned to the Hox -CO state. The spectroscopic features observed are similar to those of the [FeFe]-hydrogenases class, but relevant differences were observed given the different protein environment hosting the H-cluster.

show abstract

Expression of different types of [FeFe]-hydrogenase genes in bacteria isolated from a population of a bio-hydrogen pilot-scale plant

Cited by 45 publications

References 70 publications

Hydrogen production at high Faradaic efficiency by a bio-electrode based on TiO2 adsorption of a new [FeFe]-hydrogenase from Clostridium perfringens

Hydrogen production at high Faradaic efficiency by a bio-electrode based on TiO2 adsorption of a new [FeFe]-hydrogenase from Clostridium perfringens

Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases

Isolation and characterization of a new [FeFe]‐hydrogenase from Clostridium perfringens

Contact Info

Product

Resources

About