Biochemical and genetic characterization of the Enterococcus faecalis oxaloacetate decarboxylase complex.

Abstract:

:Enterococcus faecalis encodes a biotin-dependent oxaloacetate decarboxylase (OAD), which is constituted by four subunits: E. faecalis carboxyltransferase subunit OadA (termed Ef-A), membrane pump Ef-B, biotin acceptor protein Ef-D, and the novel subunit Ef-H. Our results show that in E. faecalis, subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex (termed Ef-AHD) which is also associated with the membrane. In order to characterize the role of the novel Ef-H subunit, coexpression of oad genes was performed in Escherichia coli, showing that this subunit is vital for Ef-A and Ef-D interaction. Diminished growth of the oadA and oadD single deletion mutants in citrate-supplemented medium indicated that the activity of the complex is essential for citrate utilization. Remarkably, the oadB-deficient strain was still capable of growing to wild-type levels but with a delay during the citrate-consuming phase, suggesting that the soluble Ef-AHD complex is functional in E. faecalis. These results suggest that the Ef-AHD complex is active in its soluble form, and that it is capable of interacting in a dynamic way with the membrane-bound Ef-B subunit to achieve its maximal alkalinization capacity during citrate fermentation.

journal_name

Appl Environ Microbiol

authors

Repizo GD,Blancato VS,Mortera P,Lolkema JS,Magni C

doi

10.1128/AEM.03980-12

subject

Has Abstract

pub_date

2013-05-01 00:00:00

pages

2882-90

issue

9

eissn

0099-2240

issn

1098-5336

pii

AEM.03980-12

journal_volume

79

pub_type

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