Efficient Expression of Haloarchaeal Nucleoside Diphosphate Kinase Via Strong Porin Promoter in Moderately Halophilic Bacteria

Chizuru      Nagayoshi; Hiroko      Tokunaga; Aya      Hayashi; Hiroaki      Harazono; Kyoko      Hamasaki; Ayumi      Ando; Masao      Tokunaga

doi:10.2174/092986606777145760

Abstract

Enzymes from extremely halophilic archaea require high concentration of salts for their proper folding and consequently are expressed as an unfolded and inactive form in Escherichia coli. Moderate halophile, which accumulates protein stabilizers, i.e., compatible solutes, is an attractive host cell for the recombinant production of heterologous proteins, since such protein stabilizers may help folding of expressed proteins. Here, we succeeded in efficient expression and purification to homogeneity of recombinant haloarchaeal nucleoside diphosphate kinase (HsNDK) in moderate halophile using newly isolated strong porin promoter.

Keywords: Halophilic, porin, protein expression, nucleoside diphosphate kinase, β-lactamase, reporter gene

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Protein & Peptide Letters

Title: Efficient Expression of Haloarchaeal Nucleoside Diphosphate Kinase Via Strong Porin Promoter in Moderately Halophilic Bacteria

Volume: 13 Issue: 6

Author(s): Chizuru Nagayoshi, Hiroko Tokunaga, Aya Hayashi, Hiroaki Harazono, Kyoko Hamasaki, Ayumi Ando and Masao Tokunaga

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Keywords: Halophilic, porin, protein expression, nucleoside diphosphate kinase, β-lactamase, reporter gene

Abstract: Enzymes from extremely halophilic archaea require high concentration of salts for their proper folding and consequently are expressed as an unfolded and inactive form in Escherichia coli. Moderate halophile, which accumulates protein stabilizers, i.e., compatible solutes, is an attractive host cell for the recombinant production of heterologous proteins, since such protein stabilizers may help folding of expressed proteins. Here, we succeeded in efficient expression and purification to homogeneity of recombinant haloarchaeal nucleoside diphosphate kinase (HsNDK) in moderate halophile using newly isolated strong porin promoter.

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Nagayoshi Chizuru, Tokunaga Hiroko, Hayashi Aya, Harazono Hiroaki, Hamasaki Kyoko, Ando Ayumi and Tokunaga Masao, Efficient Expression of Haloarchaeal Nucleoside Diphosphate Kinase Via Strong Porin Promoter in Moderately Halophilic Bacteria, Protein & Peptide Letters 2006; 13 (6) . https://dx.doi.org/10.2174/092986606777145760