TY - JOUR
T1 - Thermostable xylanases, Xyn10A and Xyn11A, from the actinomycete Nonomuraea flexuosa
T2 - solation of the genes and characterization of recombinant Xyn11A polypeptides produced in Trichoderma reesei
AU - Leskinen, S.
AU - Mäntylä, Arja
AU - Fagerström, Richard
AU - Vehmaanperä, Jari
AU - Lantto, Raija
AU - Paloheimo, Marja
AU - Suominen, Pirkko
PY - 2005
Y1 - 2005
N2 - Two endoxylanases, Nf Xyn11A and Nf Xyn10A, were cloned from a Nonomuraea flexuosa (previously Actinomadura flexuosa) DSM43186 genomic expression library in Escherichia coli. The coding sequences of xyn11A and xyn10A
consist of 344 and 492 amino acids, respectively. The catalytic domains
belong to family 11 and family 10 of glycoside hydrolases. The
C-termini share strong amino acid sequence similarity to
carbohydrate-binding module (CBM) families CBM2 and CBM13, respectively.
Native Nf Xyn11A, and recombinant Xyn11A expressed in the filamentous
fungus Trichoderma reesei, were
purified from cultivation media and characterized. The molecular masses
of the full-length enzymes determined by mass spectrometry were 32.9 kDa
and 33.4 kDa, the recombinant enzyme having higher molecular mass due
to glycosylation. In addition, shorter polypeptides with molecular
masses of 23.8 kDa and 22.0 kDa were characterized from the T. reesei
culture medium, both lacking the C-terminal CBM and the 22.0 kDa
polypeptide also lacking most of the linker region. The recombinant
polypeptides were similar to each other in terms of specific activity,
pH and temperature dependence. However, the 23.8 kDa and 22.0 kDa
polypeptides were more thermostable at 80°C than the full-length enzyme.
All polypeptide forms were effective in pretreatment of softwood kraft
pulp at 80°C.
AB - Two endoxylanases, Nf Xyn11A and Nf Xyn10A, were cloned from a Nonomuraea flexuosa (previously Actinomadura flexuosa) DSM43186 genomic expression library in Escherichia coli. The coding sequences of xyn11A and xyn10A
consist of 344 and 492 amino acids, respectively. The catalytic domains
belong to family 11 and family 10 of glycoside hydrolases. The
C-termini share strong amino acid sequence similarity to
carbohydrate-binding module (CBM) families CBM2 and CBM13, respectively.
Native Nf Xyn11A, and recombinant Xyn11A expressed in the filamentous
fungus Trichoderma reesei, were
purified from cultivation media and characterized. The molecular masses
of the full-length enzymes determined by mass spectrometry were 32.9 kDa
and 33.4 kDa, the recombinant enzyme having higher molecular mass due
to glycosylation. In addition, shorter polypeptides with molecular
masses of 23.8 kDa and 22.0 kDa were characterized from the T. reesei
culture medium, both lacking the C-terminal CBM and the 22.0 kDa
polypeptide also lacking most of the linker region. The recombinant
polypeptides were similar to each other in terms of specific activity,
pH and temperature dependence. However, the 23.8 kDa and 22.0 kDa
polypeptides were more thermostable at 80°C than the full-length enzyme.
All polypeptide forms were effective in pretreatment of softwood kraft
pulp at 80°C.
U2 - 10.1007/s00253-004-1797-x
DO - 10.1007/s00253-004-1797-x
M3 - Article
SN - 0175-7598
VL - 67
SP - 495
EP - 505
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
IS - 4
ER -