Purification and Characterization of Two Alkaline, Thermotolerant α-Amylases from Bacillus halodurans 38C-2-1 and Expression of the Cloned Gene in Escherichia coli
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概要
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A newly isolated strain, 38C-2-1, produced alkaline and thermotolerant α-amylases and was identified as Bacillus halodurans. The enzymes were purified to homogeneity and named α-amylase I and II. These showed molecular masses of 105 and 75 kDa respectively and showed maximal activities at 50–60 °C and pH 10–11, and 42 and 38% relative activities at 30 °C. These results indicate that the enzymes are thermotolerant. The enzyme activity was not inhibited by a surfactant or a bleaching reagent used in detergents. A gene encoding α-amylase I was cloned and named amyI. Production of AmyI with a signal peptide repressed the growth of an Escherichia coli transformant. When enzyme production was induced by the addition of isopropyl β-D(−)-thiogalactopyranoside in the late exponential growth phase, the highest enzyme yield was observed. It was 45-fold that of the parent strain 38C-2-1.
- 社団法人 日本農芸化学会の論文
著者
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Takenaka Shinji
Laboratory of Applied Microbiology, Department of Biofunctional Chemistry, Faculty of Agriculture, K
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Murakami Shuichiro
Laboratory of Applied Microbiology, Department of Biofunctional Chemistry, Faculty of Agriculture, K
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Aoki Kenji
Laboratory of Applied Microbiology, Department of Biofunctional Chemistry, Faculty of Agriculture, K
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Limpaseni Tipaporn
Cyclodextrin And Starch Research Unit Department Of Biochemistry Faculty Of Science Chulalongkorn Un
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Murakami Shuichiro
Dep. Of Bioscience Biotechnology And Agrochemistry Fac. Of Agriculture Meiji Univ.
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NISHIMOTO Haruka
Laboratory of Applied Microbiology, Department of Agrobioscience, Graduate School of Agricultural Sc
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TOYAMA Yosuke
Laboratory of Applied Microbiology, Department of Agrobioscience, Graduate School of Agricultural Sc
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SHIMAMOTO Etsuko
Laboratory of Applied Microbiology, Department of Agrobioscience, Graduate School of Agricultural Sc
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KAULPIBOON Jarunee
Department of Pre-Clinical Science (Biochemistry), Faculty of Medicine, Thammasat University
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PROUSOONTORN Manchumas
Cyclodextrin and Starch Research Unit, Department of Biochemistry, Faculty of Science, Chulalongkorn
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PONGSAWASDI Piamsook
Cyclodextrin and Starch Research Unit, Department of Biochemistry, Faculty of Science, Chulalongkorn
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