Using gas chromatography/isotope ratio mass spectrometry to determine the fractionation factor for H2 production by hydrogenases.
Rapid Commun Mass Spectrom
; 26(1): 61-8, 2012 Jan 15.
Article
em En
| MEDLINE
| ID: mdl-22215579
ABSTRACT
Hydrogenases catalyze the reversible formation of H(2), and they are key enzymes in the biological cycling of H(2). H isotopes have the potential to be a very useful tool in quantifying hydrogen ion trafficking in biological H(2) production processes, but there are several obstacles that have thus far limited the application of this tool. Here, we describe a new method that overcomes some of these barriers and is specifically designed to measure isotopic fractionation during enzyme-catalyzed H(2) evolution. A key feature of this technique is that purified hydrogenases are employed, allowing precise control over the reaction conditions and therefore a high level of precision. In addition, a custom-designed high-throughput gas chromatograph/isotope ratio mass spectrometer is employed to measure the isotope ratio of the H(2). Using our new approach, we determined that the fractionation factor for H(2) production by the [NiFe]-hydrogenase from Desulfovibrio fructosovorans is 0.273 ± 0.006. This result indicates that, as expected, protons are highly favored over deuterium ions during H(2) evolution. Potential applications of this newly developed method are discussed.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Hidrogênio
/
Hidrogenase
/
Cromatografia Gasosa-Espectrometria de Massas
Idioma:
En
Revista:
Rapid Commun Mass Spectrom
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Estados Unidos