Molecular cloning and expression of a novel trehalose synthase gene from <it>Enterobacter hormaechei</it>

<p>Abstract</p> <p>Background</p> <p>Trehalose synthase (TreS) which converts maltose to trehalose is considered to be a potential biocatalyst for trehalose production. This enzymatic process has the advantage of simple reaction and employs an inexpensive substrate. The...

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Main Authors: Yue Ming, Wu Xiu, Gong Wei, Ding Hong
Format: Article
Language:English
Published: BMC 2009-06-01
Series:Microbial Cell Factories
Online Access:http://www.microbialcellfactories.com/content/8/1/34
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spelling doaj-2ac582391ea7410e86fc787d062e5ae72020-11-25T00:37:10ZengBMCMicrobial Cell Factories1475-28592009-06-01813410.1186/1475-2859-8-34Molecular cloning and expression of a novel trehalose synthase gene from <it>Enterobacter hormaechei</it>Yue MingWu XiuGong WeiDing Hong<p>Abstract</p> <p>Background</p> <p>Trehalose synthase (TreS) which converts maltose to trehalose is considered to be a potential biocatalyst for trehalose production. This enzymatic process has the advantage of simple reaction and employs an inexpensive substrate. Therefore, new TreS producing bacteria with suitable enzyme properties are expected to be isolated from extreme environment.</p> <p>Results</p> <p>Six TreS producing strains were isolated from a specimen obtained from soil of the Tibetan Plateau using degenerate PCR. A novel <it>treS </it>gene from <it>Enterobacter hormaechei </it>was amplified using thermal asymmetric interlaced PCR. The gene contained a 1626 bp open reading frame encoding 541 amino acids. The gene was expressed in <it>Escherichia coli</it>, and the recombinant TreS was purified and characterized. The purified TreS had a molecular mass of 65 kDa and an activity of 18.5 U/mg. The optimum temperature and pH for the converting reaction were 37°C and 6, respectively. Hg<sup>2+</sup>, Zn<sup>2+</sup>, Cu<sup>2+</sup>and SDS inhibited the enzyme activity at different levels whereas Mn<sup>2+ </sup>showed an enhancing effect by 10%.</p> <p>Conclusion</p> <p>In this study, several TreS producing strains were screened from a source of soil bacteria. The characterization of the recombinant TreS of <it>Enterobacter hormaechei </it>suggested its potential application. Consequently, a strategy for isolation of TreS producing strains and cloning of novel <it>treS </it>genes from natural sources was demonstrated.</p> http://www.microbialcellfactories.com/content/8/1/34
collection DOAJ
language English
format Article
sources DOAJ
author Yue Ming
Wu Xiu
Gong Wei
Ding Hong
spellingShingle Yue Ming
Wu Xiu
Gong Wei
Ding Hong
Molecular cloning and expression of a novel trehalose synthase gene from <it>Enterobacter hormaechei</it>
Microbial Cell Factories
author_facet Yue Ming
Wu Xiu
Gong Wei
Ding Hong
author_sort Yue Ming
title Molecular cloning and expression of a novel trehalose synthase gene from <it>Enterobacter hormaechei</it>
title_short Molecular cloning and expression of a novel trehalose synthase gene from <it>Enterobacter hormaechei</it>
title_full Molecular cloning and expression of a novel trehalose synthase gene from <it>Enterobacter hormaechei</it>
title_fullStr Molecular cloning and expression of a novel trehalose synthase gene from <it>Enterobacter hormaechei</it>
title_full_unstemmed Molecular cloning and expression of a novel trehalose synthase gene from <it>Enterobacter hormaechei</it>
title_sort molecular cloning and expression of a novel trehalose synthase gene from <it>enterobacter hormaechei</it>
publisher BMC
series Microbial Cell Factories
issn 1475-2859
publishDate 2009-06-01
description <p>Abstract</p> <p>Background</p> <p>Trehalose synthase (TreS) which converts maltose to trehalose is considered to be a potential biocatalyst for trehalose production. This enzymatic process has the advantage of simple reaction and employs an inexpensive substrate. Therefore, new TreS producing bacteria with suitable enzyme properties are expected to be isolated from extreme environment.</p> <p>Results</p> <p>Six TreS producing strains were isolated from a specimen obtained from soil of the Tibetan Plateau using degenerate PCR. A novel <it>treS </it>gene from <it>Enterobacter hormaechei </it>was amplified using thermal asymmetric interlaced PCR. The gene contained a 1626 bp open reading frame encoding 541 amino acids. The gene was expressed in <it>Escherichia coli</it>, and the recombinant TreS was purified and characterized. The purified TreS had a molecular mass of 65 kDa and an activity of 18.5 U/mg. The optimum temperature and pH for the converting reaction were 37°C and 6, respectively. Hg<sup>2+</sup>, Zn<sup>2+</sup>, Cu<sup>2+</sup>and SDS inhibited the enzyme activity at different levels whereas Mn<sup>2+ </sup>showed an enhancing effect by 10%.</p> <p>Conclusion</p> <p>In this study, several TreS producing strains were screened from a source of soil bacteria. The characterization of the recombinant TreS of <it>Enterobacter hormaechei </it>suggested its potential application. Consequently, a strategy for isolation of TreS producing strains and cloning of novel <it>treS </it>genes from natural sources was demonstrated.</p>
url http://www.microbialcellfactories.com/content/8/1/34
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