title: Distinct actions by Paenibacillus sp. strain E18 α-L-arabinofuranosidases and xylanase in xylan degradation.
abstract: We cloned a Paenibacillus sp. strain E18 5.3-kb xylanolytic gene cluster that contains three open reading frames encoding two family 43 α-L-arabinofuranosidases (Abf43A and Abf43B) and one family 10 xylanase (XynBE18). The deduced amino acid sequences of Abf43A and Abf43B were at most 68% and 63% identical to those of two putative family 43 proteins from Clostridium sp. strain DL-VIII (EHI98634.1 and EHI98635.1), respectively, but were only 11% identical to each other. Recombinant Abf43A and Abf43B had similar activities at 45°C and pH 6.0 but varied in thermostabilities and substrate specificities. Abf43B was active against only 4-nitrophenyl α-L-arabinofuranoside, whereas Abf43A acted on 4-nitrophenyl α-L-arabinofuranoside, wheat arabinoxylan, 4-nitrophenyl α-D-xylopyranoside, and sugar beet arabinan. The sequential and combined effects on xylan degradation by XynBE18, Abf43A, and Abf43B were characterized. For beechwood, birchwood, and oat spelt xylans as the substrates, synergistic effects were found when XynBE18 and Abf43A or Abf43B were incubated together and when the substrates were first incubated with Abf43A or Abf43B and then with XynBE18. Further high-performance liquid chromatography (HPLC) analysis showed that the amounts of xylobiose and xylose increased sharply in the aforementioned reactions. For water-soluble wheat arabinoxylan as the substrate, Abf43A not only released arabinose but also had a synergistic effect with XynBE18. Synergy may arise as the result of removal of arabinose residues from xylans by α-L-arabinofuranosidases, which eliminates steric hindrance caused by the arabinose side chains and which allows xylanases to then degrade the xylan backbone, producing short xylooligosaccharides.
authors: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, People's Republic of China.
authors: Pengjun
authors: P
authors: Shi
authors: Xiaoyan
authors: X
authors: Chen
authors: Kun
authors: K
authors: Meng
authors: Huoqing
authors: H
authors: Huang
authors: Yingguo
authors: Y
authors: Bai
authors: Huiying
authors: H
authors: Luo
authors: Peilong
authors: P
authors: Yang
authors: Bin
authors: B
authors: Yao
mesh_terms: D001089:Arabinose / Q000378:metabolism; D018554:Avena / Q000737:chemistry; D029662:Betula / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004187:Disaccharides / Q000378:metabolism; D004795:Enzyme Stability; D029964:Fagus / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D056507:Paenibacillus / Q000201:enzymology*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature; D014934:Wood / Q000737:chemistry; D014990:Xylans / Q000302:isolation & purification / Q000378:metabolism*; D014994:Xylose / Q000378:metabolism; D014995:Xylosidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial; D004187:Disaccharides; D014990:Xylans; D014994:Xylose; D001089:Arabinose; D006026:Glycoside Hydrolases; D014995:Xylosidases; C019713:alpha-N-arabinofuranosidase; C004173:xylobiose