Cloning, Expression, and Transcription Analysis of L-Arabinose Isomerase Gene from Mycobacterium smegmatis SMDU
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概要
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The L-arabinose metabolic gene cluster, araA, araB, araD, araG, araH and araR, encoding L-arabinose isomerase (L-AI) and its accessory proteins was cloned from Mycobacterium smegmatis SMDU and sequenced. The deduced amino acid sequence of araA displayed highest identity with that of Bacillus subtilis (52%). These six genes comprised the L-arabinose operon, and its genetic arrangement was similar to that of B. subtilis. The L-AI gene (araA), encoding a 501 amino acid protein with a calculated molecular mass of 54,888 Da, was expressed in Escherichia coli. The productivity and overall enzymatic properties of the recombinant L-AI were almost same as the authentic L-AI from M. smegmatis. Although the recombinant L-AI showed high substrate specificity, as did L-AI from other organisms, this enzyme catalyzed not only isomerization of L-arabinose-L-ribulose and D-galactose-D-tagatose but also isomerization of L-altrose-L-psicose and L-erythrulose-L-threose. In combination with L-AI from M. smegmatis, L-threose and L-altrose can be produced from cheap and abundant erythritol and D-fructose respectively, indicating that this enzyme has great potential for biological application in rare sugar production. Transcription analysis using various sugars revealed that this enzyme was significantly induced not only by L-arabinose and D-galactose but also by L-ribose, galactitol, L-ribulose, and L-talitol. This different result of transcription mediated by sugars from that of E. coli suggests that the transcriptional regulation of araA from M. smegmatis against sugar is loose compared with that from E. coli, and that it depends on the hydroxyl configuration at C2, C3 and C4 positions of sugars.
- 社団法人 日本農芸化学会の論文
著者
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Izumori Ken
Rare Sugar Research Center Faculty Of Agriculture Kagawa University
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Takata Goro
Rare Sugar Res. Center And Fac. Of Agriculture Kagawa Univeristy
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Takada Goro
Department Of Biochemistry And Food Science Faculty Of Agriculture Kanagawa University
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Morimoto Kenji
Rare Sugar Research Center Kagawa University
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Rao Devendar
Rare Sugar Research Center Faculty Of Agriculture Kagawa University
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Poonperm Wayoon
Rare Sugar Research Center, Kagawa University
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SOUDA Akane
Rare Sugar Research Center, Kagawa University
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NISHIZAKI Tomoe
Rare Sugar Research Center, Kagawa University
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