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Molecular Cloning of the Gene Encoding Thermostable Endo-1,5-α-L-arabinase ofBacillus thermodenitrificansTS-3 and Its Expression inBacillus subtilis
Author(s) -
Makoto Takao,
Asako Yamaguchi,
Kentaro Yoshikawa,
Takao Terashita,
Takuo Sakai
Publication year - 2002
Publication title -
bioscience biotechnology and biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.509
H-Index - 116
eISSN - 1347-6947
pISSN - 0916-8451
DOI - 10.1271/bbb.66.430
Subject(s) - bacillus subtilis , gene , open reading frame , amino acid , biology , cloning (programming) , pseudomonas fluorescens , molecular cloning , peptide sequence , biochemistry , genetics , bacteria , computer science , programming language
The gene that encodes a thermostable endo-arabinase (called ABN-TS) from Bacillus thermodenitrificans TS-3 was cloned, sequenced, and expressed in the mesophilic B. subtilis. The gene contained an open reading frame consists of 939 bp, which encodes 313 amino acids. The deduced amino acid sequence of the enzyme showed 50, 46, and 36% similarity with endo-arabinase from B. subtilis IFO 3134 (PPase-C), Pseudomonas fluorescens (ArbA), and Aspergillus niger (ABNA), respectively. The hydrophobic and acidic amino acids making up ABN-TS outnumbered those in PPase-C. The gene product expressed in B. subtilis, as the host, had substantially the same characteristics, and was stable up to 70 degrees C, and the reaction was optimal around 70 degrees C, as well as native ABN-TS.

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