Gene cloning, expression, and characterization of a family 51 alpha-L-arabinofuranosidase from Streptomyces sp. S9.
An alpha-L-arabinofuranosidase gene, abf51S9, was cloned from Streptomyces sp. S9 and successfully expressed in Escherichia coli BL21 (DE3). The full-length gene consisted of 1,506 bp and encoded 501 amino acids with a calculated mass of 55.2 kDa. The deduced amino acid sequence was highly homologous with the alpha-L-arabinofuranosidases belonging to family 51 of the glycoside hydrolases. The recombinant protein was purified to electrophoretic homogeneity by Ni-NTA affinity chromatography and subsequently characterized. The optimal pH and temperature for the recombinant enzyme were 6.0 and 60 approximately 65 degrees C, respectively. The enzyme showed a broad pH range of stability, retaining over 75% of the maximum activity at pH 5.0 to 11.0. The specific activity, K (m), and V (max) with p-nitrophenyl-alpha-L-arabinofuranoside as substrate were 60.0 U mg(-1), 1.45 mM, and 221 mumol min(-1) mg(-1), respectively. Abf51S9 showed a mild but significant synergistic effect in combination with xylanase on the degradation of oat-spelt xylan and soluble wheat arabinoxylan substrates with a 1.19- and 1.21-fold increase in the amount of reducing sugar released, respectively. These favorable properties make Abf51S9 a good candidate in various industrial applications.
Shi
Pengjun
P
Key Laboratory for Feed Biotechnology of Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China.
Li
Ning
N
Yang
Peilong
P
Wang
Yaru
Y
Luo
Huiying
H
Bai
Yingguo
Y
Yao
Bin
B
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology*; D014990:Xylans / Q000378:metabolism
D001426:Bacterial Proteins; D014990:Xylans; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase