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Agrobacterium tumefaciens type II NADH dehydrogenase
Author(s) -
Bernard Laetitia,
Desplats Carine,
Mus Florence,
Cuiné Stéphan,
Cournac Laurent,
Peltier Gilles
Publication year - 2006
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2006.05370.x
Subject(s) - agrobacterium tumefaciens , dehydrogenase , nadh dehydrogenase , biology , microbiology and biotechnology , enzyme , biochemistry , transformation (genetics) , gene , protein subunit
Type II NADH dehydrogenases (NDH‐2) are monomeric enzymes that catalyse quinone reduction and allow electrons to enter the respiratory chain in different organisms including higher plant mitochondria, bacteria and yeasts. In this study, an Agrobacterium tumefaciens gene encoding a putative alternative NADH dehydrogenase (AtuNDH‐2) was isolated and expressed in Escherichia coli as a (His) 6 ‐tagged protein. The purified 46 kDa protein contains FAD as a prosthetic group and oxidizes both NADH and NADPH with similar V max values, but with a much higher affinity for NADH than for NADPH. AtuNDH‐2 complements the growth (on a minimal medium) of an E. coli mutant strain deficient in both NDH‐1 and NDH‐2, and is shown to supply electrons to the respiratory chain when incubated with bacterial membranes prepared from this mutant. By measuring photosystem II chlorophyll fluorescence on thylakoid membranes prepared from the green alga Chlamydomonas reinhardtii , we show that AtuNDH‐2 is able to stimulate NADH‐dependent reduction of the plastoquinone pool. We discuss the possibility of using heterologous expression of NDH‐2 enzymes to improve nonphotochemical reduction of plastoquinones and H 2 production in C. reinhardtii .