The nasFEDCBA operon for nitrate and nitrite assimilation in Klebsiella pneumoniae M5al.

Abstract

Klebsiella pneumoniae can use nitrate and nitrite as sole nitrogen sources through the nitrate assimilation pathway. We previously identified structural genes for assimilatory nitrate and nitrite reductases, nasA and nasB, respectively. We report here our further identification of four genes, nasFEDC, upstream of the nasBA genes. The nasFEDCBA genes probably form an operon. Mutational and complementation analyses indicated that both the nasC and nasA genes are required for nitrate assimilation. The predicted NASC protein is homologous to a variety of NADH-dependent oxidoreductases. Thus, the NASC protein probably mediates electron transfer from NADH to the NASA protein, which contains the active site for nitrate reduction. The deduced NASF, NASE, and NASD proteins are homologous to the NRTA, NRTB, and NRTD proteins, respectively, that are involved in nitrate uptake in Synechococcus sp. (T. Omata, X. Andriesse, and A. Hirano, Mol. Gen. Genet. 236:193-202, 1993). Mutational and complementation studies indicated that the nasD gene is required for nitrate but not nitrite assimilation. By analogy with the Synechococcus nrt genes, we propose that the nasFED genes are involved in nitrate transport in K. pneumoniae.