Ureases of Extreme Halophiles of the Genus Haloarcula with a Unique Structure of Gene Cluster
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概要
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We searched for urease activities in 71 strains of extreme halophiles by a urea-phenol red-agar plate method. Positive strains were further investigated by measuring the ammonia released from urea in cell-free extracts. Only 4 strains of the genus Haloarcula, Har. aidinensis, Har. hispanica, Har. japonica, and Har. marismortui were finally shown as the urease producers. A partially purified urease from Har. hispanica was a typical halophilic enzyme in that it showed maximum activity at 18–23% NaCl and lost the activity irreversibly in the absence of NaCl. Partial genes (1596 bp) of the urease encoding from upstream of the β subunit down to the N-terminal 139 amino acids of the α subunit, were PCR amplified from the four strains, as well as from five urease-negative Haloarcula strains. Strains of other genera, which were urease-negative, did not yield PCR products. The deduced amino acid sequences of the β subunit and partial α subunit were similar to each other (92–100% similarities) and to those from other organisms. Analysis of the draft genome sequence of Har. marismortui, however, suggested that the order of the genes encoding the three subunits (with the total number of amino acids of 834) and four accessory proteins was β-α-γ-UreG-UreD-UreE-UreF. This order is quite unique, since in other microorganisms the order is γ-β-α-UreE-UreF-UreG-UreD in most cases. No open reading frames were detected in the PCR-amplified upstream of the β subunit, suggesting that all Haloarcula species have the same unique structure of the urease gene cluster.
- 社団法人 日本農芸化学会の論文
- 2004-02-23
著者
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HORIKOSHI Koki
Department of Bioengineering, The Tokyo Institute of Technology
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Kawano H
Extremobiosphere Research Center (xbr) Japan Agency For Marine-earth Science And Technology (jamstec
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Usami R
Department Of Applied Chemistry Faculty Of Engineering Toyo University
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Usami R
Toyo Univ. Saitama Jpn
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Usami Ron
Department Of Applied Chemistry Faculty Of Engineering Toyo University
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Horikoshi K
Extremobiosphere Res. Center (xbr) Japan Agency For Marine-earth Sci. And Technol. (jamstec)
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Yoshida Yasuhiko
Department Of Applied Chemistry Faculty Of Engineering Toyo University
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KAMEKURA Masahiro
Noda Institute for Scientific Research
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Yoshida Y
Department Of Applied Chemistry Faculty Of Engineering Toyo University:bio-nano Electronics Research
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Usami Ron
Graduate School Of Interdisciplinary New Science Toyo University
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FUKUSHIMA TADAMASA
Department of Applied Chemistry, Faculty of Engineering, Toyo University
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MIZUKI TORU
Department of Applied Chemistry, Faculty of Engineering, Toyo University
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DASSARMA Shiladitya
Center for Marine Biotechnology, University of Maryland Biotechnology Institute
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Horikoshi Koki
Department Of Applied Chemistry Faculty Of Engineering Toyo University
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Mizuki Toru
Department Of Applied Chemistry Faculty Of Engineering Toyo University
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Dassarma Shiladitya
Center For Marine Biotechnology University Of Maryland Biotechnology Institute
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Fukushima Tadamasa
Department Of Applied Chemistry Faculty Of Engineering Toyo University:bio-nano Electronics Research
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