The C-terminal region of xylanase domain in Xyn11A from Paenibacillus curdlanolyticus B-6 plays an important role in structural stability

บทความในวารสาร

ผู้เขียน/บรรณาธิการ

กลุ่มสาขาการวิจัยเชิงกลยุทธ์

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รายละเอียดสำหรับงานพิมพ์

รายชื่อผู้แต่ง: Sermsathanaswadi J., Pianwanit S., Pason P., Waeonukul R., Tachaapaikoon C., Ratanakhanokchai K., Septiningrum K., Kosugi A.

ผู้เผยแพร่: Springer

ปีที่เผยแพร่ (ค.ศ.): 2014

วารสาร: Applied Microbiology and Biotechnology (0175-7598)

Volume number: 98

Issue number: 19

หน้าแรก: 8223

หน้าสุดท้าย: 8233

จำนวนหน้า: 11

นอก: 0175-7598

eISSN: 1432-0614

URL: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84920253530&doi=10.1007%2fs00253-014-5748-x&partnerID=40&md5=18c82ad45067a4ba6e93aa07c3ed28d7

ภาษา: English-Great Britain (EN-GB)

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บทคัดย่อ

Paenibacillus curdlanolyticus B-6 produces an extracellular multienzyme complex containing a major xylanase subunit, designated Xyn11A, which includes two functional domains belonging to glycosyl hydrolase family-11 (GH11) and carbohydrate binding module family-36 (CBM36) and possesses a glycine and asparagine-rich linker (linker). To clarify the roles of each functional domain, recombinant proteins XynXL and XynX (CBM36 deleted and CBM36 and linker deleted, respectively) were constructed. Their xylanase activities were similar toward soluble xylan, whereas XynXL showed decreased hydrolysis activity toward insoluble xylan while XynX had no xylanase activity. To determine the significance of the linker and its neighbor region, XynX was subjected to secondary structural alignments using circular dichroism (CD) spectroscopy and three-dimensional (3D) structural analysis. A seven amino acid (NTITIGG) neighbor linker sequence was highly conserved among GH11 xylanases of Paenibacillus species. Although XynX exhibited a typical GH11 xylanase structure, conformational gaps were observed in the β6- and β12-sheets and in CD spectra. Flipping of the Arg163 side chains in the subsite was also observed upon analysis of superimposed models. Docking analysis using xylohexaose indicated that flipping of the Arg163 side chains markedly affected substrate binding in the subsite. To identify the amino acids related to stabilizing the substrate binding site, XynX with an extended C-terminal region was designed. At least seven amino acids were necessary to recover substrate binding and xylanase activity. These results indicated that the seven amino acid neighbor Xyn11A linker plays an important role in the activity and conformational stability of the xylanase domain. © 2014, Springer-Verlag Berlin Heidelberg.

คำสำคัญ

CBM, GH family-11