Purification and characterization of a thermostable xylanase from Paenibacillus sp. NF1 and its application in xylooligosaccharides production.
High levels of extracellular xylanase activity (211.79 IU/mg) produced by Paenibacillus sp. NF1 were detected when it was submerged-cultured. After three consecutive purification steps using Octyl-Sepharose, Sephadex G75, and Q-Sepharose columns, a thermostable xylanase (XynNF) was purified to homogeneity and showed a molecular mass of 37 kDa according to SDS-PAGE. The specific activity of the purified XynNF was up to 3,081.05 IU/mg with a 14.55-fold purification. The activity of XynNF was stimulated by Ca(2+), Ba(2+), DTT, and β-mercaptoethanol, but was inhibited by Fe(3+), Zn(2+), Fe(2+), Cu(2+), SDS, and EDTA. The purified XynNF displayed a greater affinity for oat spelt xylan with the maximal enzymatic activity at 60°C and pH 6.0. XynNF, which was shown to be cellulose-free, with high stability at high temperature (70°C-80°C) and low pH range (pH 4.0-7.0), is potentially valuable for various industrial applications. The end products of high efficient oat spelt xylan hydrolysis by XynNF (an endoxylanase) containing 95.8% xylooligosaccharides of 2-4 degree of polymerization (DP2-4) with the enrichment of xylobiose (61.5%) indicated that XynNF is a promising candidate for xylooligosaccharides production.
Chinese Academy of Agricultural Engineering, 100125 Beijing, China.
Hong-chen
HC
Zheng
Ming-zhe
MZ
Sun
Ling-cai
LC
Meng
Hai-sheng
HS
Pei
Xiu-qing
XQ
Zhang
Zheng
Z
Yan
Wen-hui
WH
Zeng
Jing-sheng
JS
Zhang
Jin-rong
JR
Hu
Fu-ping
FP
Lu
Jun-she
JS
Sun
D018554:Avena / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D020536:Enzyme Activators / Q000032:analysis; D004791:Enzyme Inhibitors / Q000032:analysis; D004795:Enzyme Stability; D005965:Glucuronates / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism*; D056507:Paenibacillus / Q000201:enzymology*; D013696:Temperature; D014990:Xylans / Q000378:metabolism; D014995:Xylosidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*
D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D005965:Glucuronates; D009844:Oligosaccharides; D014990:Xylans; C570991:xylooligosaccharide; D014995:Xylosidases