Cloning, expression and characterization of a trehalose synthase gene from rhodococcus opacus

文献类型: 外文期刊

第一作者: Yan, Junyan

作者: Yan, Junyan;Qiao, Yu;Ding, Hongbiao;Hu, Jun;Ding, Hongbiao

作者机构:

关键词: Escherichia coli;Rhodococcus opacus;Trehalose;Trehalose synthase

期刊名称:PROTEIN JOURNAL ( 影响因子:2.371; 五年影响因子:1.929 )

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收录情况: SCI

摘要: Trehalose is a unique disaccharide capable of protecting proteins against environmental stress. A novel trehalose synthase (TreS) gene from Rhodococcus opacus was cloned and expressed in Escherichia coli Top10 and BL21 (DE3) pLysS, respectively. The recombinant TreS showed a molecular mass of 79 kDa. Thin layer chromatography (TLC) result suggested that this enzyme had the ability to catalyze the mutual conversion of maltose and trehalose. Moreover, high-performance liquid chromatography (HPLC) result suggested that glucose appeared as a byproduct with a conversion rate of 12 %. The purified recombinant enzyme had an optimum temperature of 25 C and pH optimum around 7.0. Kinetic analysis revealed that the K m for trehalose was around 98 mM, which was a little higher than that of maltose. The preferred substrate of TreS was maltose according to the analysis of k cat/K m. Both 1 and 10 mM of Hg2+, Cu2+ and Al3+ could inhibit the TreS activity, while only 1 mM of Ca2+ and Mn2+ could increase its activity. Five amino acid residues, Asp244, Glu 286, Asp354, His147 and His353, were shown to be conserved in R. opacus TreS, which were also important for α-amylase family enzyme catalysis.

分类号: Q51

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