Effects of dockerin domains on Neocallimastix frontalis xylanases.
Two xylanase genes were cloned from the anaerobic fungus Neocallimastix frontalis. Xyn11A had a modular structure of two catalytic domains and two dockerin domains, while Xyn11B had one catalytic domain and two dockerin domains. The characteristics of the xylanases with and without dockerin domains were investigated. The deletion of dockerin domains had little influence on the optimal pH of xylanases, while it significantly affected the optimal temperatures. The optimal temperatures increased from 55 to 60 degrees C for Xyn11A and 60 to 65 degrees C for Xyn11B after the deletion of dockerin domains. The increase of optimal temperatures was attributed to the lower stability of the second structure in full length xylanase than that in the truncated one as evidenced by the circular dichroism spectroscopy. The specific activity of Xyn11A and Xyn11B increased about 64% and 330%, respectively, after the deletion of the dockerin domains. The removal of dockerin domains appeared to increase the overall efficiency of Xyn11A' (1.2-) and Xyn11B' (2.9-) fold with oat spelts xylan as reflected by the values of k(cat)/K(m). The results suggest that the dockerin domain might play an important role in the characteristics of xylanases from anaerobic fungi.
Huang
Ya-Hui
YH
Institute of Microbiology and Biochemistry, National Taiwan University, Taipei 106, Taiwan, ROC.
Huang
Ching-Tsan
CT
Hseu
Ruey-Shyang
RS
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D020088:Neocallimastix / Q000201:enzymology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D013379:Substrate Specificity; D013696:Temperature; D014990:Xylans / Q000378:metabolism*; D014995:Xylosidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001617:beta-Glucosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D014990:Xylans; D014995:Xylosidases; D001617:beta-Glucosidase; D043364:Endo-1,4-beta Xylanases; C090001:XynB xylanase