Isolation and Characterization of a Novel Thermostable Neopullulanase-Like Enzyme from a Hot Spring in Thailand
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概要
- 論文の詳細を見る
A gene encoding a thermostable pullulan-hydrolyzing enzyme was isolated from environmental genomic DNA extracted from soil sediments of Bor Khleung hot spring in Thailand. Sequence comparison with related enzymes suggested that the isolated enzyme, designated Env Npu193A, was most likely a neopullulanase-like enzyme. Env Npu193A was expressed in Pichia pastoris as a monomeric recombinant protein. The purified Env Npu193A exhibited pH stability ranging from 3 to 9. More than 60% of enzyme activity was retained after incubation at 60 °C for 1 h. Env Npu193A was found to hydrolyze various substrates, including pullulan, starch, and γ-cyclodextrin. The optimal working condition for Env Npu193A was at pH 7 at 75 °C with Km and Vmax toward pullulan of 1.22±0.3% and 23.24±1.7 U/mg respectively. Env Npu193A exhibited distinct biochemical characteristics as compared with the previously isolated enzyme from the same source. Thus, a culture-independent approach with sequence-basing was found to be an effective way to discover novel enzymes displaying unique substrate specificity and high thermostability from natural bioresources.
- 社団法人 日本農芸化学会の論文
- 2008-06-23
著者
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Sriprang Rutchadaporn
Biotec Central Research Unit National Center For Genetic Engineering And Biotechnology National Scie
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EURWILAICHITR Lily
BIOTEC Training Center of Genetic Engineering and Biotechnology at Institute of Molecular Biology an
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Champreda Verawat
Enzyme Technology Laboratory National Center For Genetic Engineering And Biotechnology
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Eurwilaichitr Lily
Enzyme Technology Laboratory National Center For Genetic Engineering And Biotechnology
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Tanapongpipat Sutipa
Enzyme Technology Laboratory Bioresources Technology Unit National Center For Genetic Engineering An
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TANG Kittapong
BIOTEC Central Research Unit, National Center for Genetic Engineering and Biotechnology, National Sc
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KOBAYASHI Rutchadaporn
BIOTEC Central Research Unit, National Center for Genetic Engineering and Biotechnology, National Sc
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CHAMPREDA Verawat
BIOTEC Central Research Unit, National Center for Genetic Engineering and Biotechnology, National Sc
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TANAPONGPIPAT Sutipa
BIOTEC Central Research Unit, National Center for Genetic Engineering and Biotechnology, National Sc
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Tang Kittapong
Biotec Central Research Unit National Center For Genetic Engineering And Biotechnology National Scie
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