Cloning and expression of an alpha-L-arabinofuranosidase gene (stxIV) from Streptomyces thermoviolaceus OPC-520, and characterization of the enzyme.
The gene encoding alpha-L-arabinofuranosidase (STX-IV), located upstream of the previously reported stxI gene, was cloned and sequenced. The gene is divergently transcribed from the stxI gene, and the two genes are separated by 661 nucleotides. The stxIV gene consists of a 1,092-bp open reading frame encoding 363 amino acids. The deduced amino acid sequence of the gene showed that STX-IV was an enzyme consisting of only a catalytic domain, and that the enzyme had significant similarity with alpha-L-arabinofuranosidases belonging to family 62 of glycosyl hydrolases. The stxIV gene was expressed in Escherichia coli, and the recombinant protein was purified to homogeneity. Arabinoxylan and oat spelt xylan were good substrates for STX-IV, however, the enzyme showed a low activity with p-nitrophenyl alpha-L-arabinofuranoside. The optimum pH and temperature were 5.0 and 60 degrees C, respectively.
Tsujibo
Hiroshi
H
Osaka University of Pharmaceutical Sciences, Takatsuki, Japan. tsujibo@gly.oups.ac.jp
Takada
Chiaki
C
Wakamatsu
Yoshiko
Y
Kosaka
Mitsuo
M
Tsuji
Akihiko
A
Miyamoto
Katsushiro
K
Inamori
Yoshihiko
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics
D004269:DNA, Bacterial; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase