Identification and characterization of a novel xylanase derived from a rice straw degrading enrichment culture.
A metagenomic library containing ca. 3.06 x 10(8) bp insert DNA was constructed from a rice straw degrading enrichment culture. A xylanase gene, umxyn10A, was cloned by screening the library for xylanase activity. The encoded enzyme Umxyn10A showed 58% identity and 73% similarity with a xylanase from Thermobifida fusca YX. Sequence analyses showed that Umxyn10A contained a glycosyl hydrolase family 10 catalytic domain. The gene was expressed in Escherichia coli, and the recombinant enzyme was purified and characterized biochemically. Recombinant Umxyn10A was highly active toward xylan. However, the purified enzyme could slightly hydrolyze beta-1,3/4-glucan and beta-1,3/6-glucan. Umxyn10A displayed maximal activity toward oat spelt xylan at a high temperature (75 degrees C) and weak acidity (pH 6.5). The K( m ) and V (max) of Umxyn10A toward oat spelt xylan were 3.2 mg ml(-1) and 0.22 mmol min(-1) mg(-1) and were 2.7 mg ml(-1) and 1.0 mmol min(-1) mg(-1) against birchwood xylan, respectively. Metal ions did not appear to be required for the catalytic activity of this enzyme. The enzyme Umxyn10A could efficiently hydrolyze birchwood xylan to release xylobiose as the major product and a negligible amount of xylose. The xylanase identified in this work may have potential application in producing xylobiose from xylan.
Mo
Xin-chun
XC
Guangxi University, Nanning, People's Republic of China.
Chen
Chun-lan
CL
Pang
Hao
H
Feng
Yi
Y
Feng
Jia-xun
JX
D000595:Amino Acid Sequence; D001419:Bacteria / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D004795:Enzyme Stability; D015870:Gene Expression; D008969:Molecular Sequence Data; D012275:Oryza / Q000382:microbiology*; D010802:Phylogeny; D016415:Sequence Alignment; D012988:Soil Microbiology; D013379:Substrate Specificity; D014990:Xylans / Q000378:metabolism
D001426:Bacterial Proteins; D014990:Xylans; D043364:Endo-1,4-beta Xylanases