Abstract
Hemicellulose represents a rich source of biomass that can be converted into useful chemical feedstocks. One of the main components of hemicellulose is xylan, a polymer of xylose residues. Xylanase enzymes that hydrolyze xylan are therefore of great commercial interest. We have cloned a gene (xyn11A) that encodes a 283-amino acid xylanase enzyme from the fungus Lentinula edodes. The enzyme has a pI of 4.6 and belongs to the highly conserved glycosyl hydrolase family 11. The xylanase gene was cloned into a Pichia pastoris expression vector that secretes active enzyme into both solid and liquid media. The optimal reaction conditions were at pH 4.5 and 50°C. The enzyme had a Km of 1.5 mg/ml and a Vmax of 2.1 mmol/min/mg. Xyn11A produced primarily xylobiose, xylotriose, and xylotetraose from a birchwood xylan substrate. This is the first report on the cloning of a hemicellulase gene from L. edodes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Asp:
-
aspartic acid
- Avi:
-
avicel
- BeX:
-
beechwood xylan
- BiX:
-
birchwood xylan
- CMC:
-
carboxymethyl cellulose
- DNSA:
-
dinitrosalicyclic acid
- OSX:
-
oat spelt xylan
- PCR:
-
polymerase chain reaction
- RACE-PCR:
-
rapid amplification of cDNA ends
- RBB:
-
remazol brilliant blue
- RFU:
-
relative fluroscence units
- VAl:
-
valine
- Xyn:
-
xylanase
- YEPD:
-
yeast extract-peptone-dextrose growth media
References
P. Bajpai (1997) Adv. Appl. Microbiol. 43 141–194
Q. K. Beg M. Kapoor L. Mahajan G. S. Hoondal (2001) Appl. Microbiol. Biotechnol. 56 326–338 Occurrence Handle10.1007/s002530100704
P. Biely C. R. MacKenzie J. Puls H. Schneider (1986) Bio/technology 4 731–733 Occurrence Handle10.1038/nbt0886-731
P. Chandrakant V. S. Bisaria (1998) Crit. Rev. Biotechnol. 18 295–331
E. Gomez-Gomez M. Isabel G. Roncero A. Di Pietro C. Hera (2001) Curr. Genet. 40 268–275 Occurrence Handle10.1007/s00294-001-0260-0
B. Henrissat A. Bairoch (1996) Biochem. J. 316 695–696
G. Johansson J. Stahlberg G. Lindeberg A. Engstrom G. Pettersson (1989) FEBS Lett. 243 389–393 Occurrence Handle10.1016/0014-5793(89)80168-1
T. Kimura H. Suzuki H. Furuhashi T. Aburatani K. Morimoto S. Karita K. Sakka K. Ohmiya (2000) Biosci. Biotechnol. Biochem. 64 2734–2738
E. P. Ko H. Akatsuka H. Moriyama A. Shinmyo Y. Hata Y. Katsube I. Urabe H. Okada (1992) Biochem. J. 288 117–121
G. F. Leatham (1985) Appl. Environ. Microbiol. 50 859–867
C. C. Lee D. W. Wong G. H. Robertson (2001) FEMS Microbiol. Lett. 205 355–360 Occurrence Handle10.1016/S0378-1097(01)00500-6
S. Miao L. Ziser R. Aebersold S. G. Withers (1994) Biochemistry 33 7027–7032
H. Nielsen J. Engelbrecht S. Brunak G. von Heijne (1997) Protein Eng. 10 1–6 Occurrence Handle10.1093/protein/10.1.1
R. A. Prade (1996) Biotechnol. Genet. Eng. Rev. 13 101–131
H. V. Tilbeurgh P. Tomme M. Claeyssens R. Bhikhabhai G. Pettersson (1986) FEBS Lett. 204 223–227 Occurrence Handle10.1016/0014-5793(86)80816-X
A. Torronen R. L. Mach R. Messner R. Gonzalez N. Kalkkinen A. Harkki C. P. Kubicek (1992) Biotechnology (N Y) 10 1461–1465 Occurrence Handle10.1038/nbt1192-1461
W. W. Wakarchuk R. L. Campbell W. L. Sung J. Davoodi M. Yaguchi (1994) Protein Sci. 3 467–475
O. P. Ward M. Moo-Young (1989) Crit. Rev. Biotechnol. 8 237–274
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, C.C., Wong, D.W.S. & Robertson, G.H. Cloning and Characterization of the Xyn11A Gene from Lentinula edodes. Protein J 24, 21–26 (2005). https://doi.org/10.1007/s10930-004-0602-0
Received:
Issue Date:
DOI: https://doi.org/10.1007/s10930-004-0602-0