The gene encoding alpha-L-arabinofuranosidase (AFase) from Paenibacillus sp. DG-22 was cloned, sequenced, and expressed in Escherichia coli. The AFase gene (abfA) comprises a 1,509 bp open reading frame encoding 502 amino acids with a molecular mass of 56,520 daltons. The deduced amino acid sequence of the gene shows that AbfA is an enzyme consisting of only a catalytic domain, and that the enzyme has significant similarity to AFases classified into the family 51 of the glycosyl hydrolases. abfA was subcloned into the pQE60 expression vector to fuse it with a six-histidine tag and the recombinant AFase (rAbfA) was purified to homogeneity. The specific activity of the recombinant enzyme was 96.7 U/mg protein. Determination of the apparent molecular mass by gel-filtration chromatography indicated that AbfA has a tetrameric structure. The optimal pH and temperature of the enzyme were 6.0 and 60 degrees C, respectively. The enzyme activity was completely inhibited by 1 mM HgCl2. rAbfA was active only towards p-nitrophephenyl alpha-L-arabinofuranoside and exhibited K-m and V-max values of 3.5 mM and 306.1 U/mg, respectively. rAbfA showed a synergistic effect in combination with endoxylanase on the degradation of oat spelt xylan and wheat arabinoxylan.
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Qujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R ChinaQujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R China
Zhu, Ling
Wang, Yuchen
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Yuxi Normal Univ, Sch Chem Biol & Environm, Yuxi 653100, Peoples R ChinaQujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R China
Wang, Yuchen
Cai, Jian
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Qujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R ChinaQujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R China
机构:
UNSAM, CONICET, IIB INTECH, Chascomus, Argentina
UNLP, Fac Ciencias Exactas, RA-1900 La Plata, ArgentinaUNSAM, CONICET, IIB INTECH, Chascomus, Argentina
Civello, Pedro M.
Martinez, Gustavo A.
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UNSAM, CONICET, IIB INTECH, Chascomus, Argentina
UNLP, Fac Ciencias Exactas, RA-1900 La Plata, ArgentinaUNSAM, CONICET, IIB INTECH, Chascomus, Argentina
机构:
Qujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R ChinaQujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R China
Zhu, Ling
Wang, Yuchen
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Yuxi Normal Univ, Sch Chem Biol & Environm, Yuxi 653100, Peoples R ChinaQujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R China
Wang, Yuchen
Cai, Jian
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Qujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R ChinaQujing Normal Univ, Yunnan Engn Res Ctr Fruit Wine, Qujing 655011, Peoples R China
机构:
UNSAM, CONICET, IIB INTECH, Chascomus, Argentina
UNLP, Fac Ciencias Exactas, RA-1900 La Plata, ArgentinaUNSAM, CONICET, IIB INTECH, Chascomus, Argentina
Civello, Pedro M.
Martinez, Gustavo A.
论文数: 0引用数: 0
h-index: 0
机构:
UNSAM, CONICET, IIB INTECH, Chascomus, Argentina
UNLP, Fac Ciencias Exactas, RA-1900 La Plata, ArgentinaUNSAM, CONICET, IIB INTECH, Chascomus, Argentina