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Can J Microbiol. 2017 Jul;63(7):596-607. doi: 10.1139/cjm-2017-0104. Epub 2017 Mar 22.

A novel NhaD-type Na+/H+ antiporter from the moderate halophile and alkaliphile Halomonas alkaliphila.

Author information

1
a Department of Microbiology and Biotechnology, College of Life Sciences, Northeast Agricultural University, Harbin, 150030, People's Republic of China.
2
b College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing, 163319, People's Republic of China.
3
c Department of Microbiology, Soils, Water & Environment Research Institute, Agriculture Research Center, Egypt.

Abstract

In this study, a NhaD-type Na+/H+ antiporter gene designated Ha-nhaD was obtained by selection of genomic DNA from the moderate halophile and alkaliphile Halomonas alkaliphila in Escherichia coli KNabc lacking 3 major Na+/H+ antiporters. The presence of Ha-NhaD conferred tolerance of E. coli KNabc to NaCl up to 0.6 mol·L-1 and to LiCl up to 0.2 mol·L-1 and to an alkaline pH. pH-dependent Na+(Li+)/H+ antiport activity was detected from everted membrane vesicles prepared from E. coli KNabc/pUC-nhaD but not those of KNabc/pUC18. Ha-NhaD exhibited Na+(Li+)/H+ antiport activity over a wide pH range from 7.0 to 9.5, with the highest activity at pH 9.0. Protein sequence alignment and phylogenetic analysis revealed that Ha-NhaD is significantly different from the 7 known NhaD-type Na+/H+ antiporters, including Dw-NhaD, Dl-NhaD, Vp-NhaD, Vc-NhaD, Aa-NhaD, He-NhaD, and Ha-NhaD1. Although Ha-NhaD showed a closer phylogenetic relationship with Ha-NhaD2, a significant difference in pH-dependent activity profile exists between Ha-NhaD and Ha-NhaD2. Taken together, Ha-nhaD encodes a novel pH-dependent NhaD-type Na+/H+ antiporter.

KEYWORDS:

Halomonas alkaliphila; Na+/H+ antiporter; NhaD; antiport Na+/H+; halophile et alcaliphile modéré; moderate halophile and alkaliphile; pH-dependent activity profile; profil d’activités dépendantes du pH

PMID:
28329448
DOI:
10.1139/cjm-2017-0104
[Indexed for MEDLINE]

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