Počet záznamů: 1
Engineering the acceptor substrate specificity in the xyloglucan endotransglycosylase TmXET6.3 from nasturtium seeds (Tropaeolum majus L.)
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SYSNO ASEP 0504319 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Engineering the acceptor substrate specificity in the xyloglucan endotransglycosylase TmXET6.3 from nasturtium seeds (Tropaeolum majus L.) Tvůrce(i) Stratilová, B. (SK)
Firáková, Z. (SK)
Klaudiny, J. (SK)
Šesták, S. (SK)
Kozmon, S. (SK)
Strouhalová, Dana (UIACH-O)
Garajová, S. (SK)
Ait‑Mohand, F. (SK)
Horváthová, A. (SK)
Farkaš, V. (SK)
Stratilová, E. (SK)
Hrmova, M. (SK)Celkový počet autorů 12 Zdroj.dok. Plant Molecular Biology. - : Springer - ISSN 0167-4412
Roč. 100, 1-2 (2019), s. 181-197Poč.str. 17 s. Forma vydání Online - E Jazyk dok. eng - angličtina Země vyd. NL - Nizozemsko Klíč. slova bioinformatics ; protein molecular modelling ; homo- and hetero-transglycosylation Vědní obor RIV CB - Analytická chemie, separace Obor OECD Analytical chemistry Vědní obor RIV – spolupráce Ústav analytické chemie - Analytická chemie, separace Způsob publikování Omezený přístup Institucionální podpora UIACH-O - RVO:68081715 UT WOS 000467940100014 EID SCOPUS 85062941288 DOI 10.1007/s11103-019-00852-8 Anotace Xyloglucan xyloglucosyl transferases (XETs) (EC 2.4.1.207) play a central role in loosening and re-arranging the cellulose-xyloglucan network, which is assumed to be the primary load-bearing structural component of plant cell walls. The sequence of mature TmXET6.3 from Tropaeolum majus (280 residues) was deduced by the nucleotide sequence analysis of complete cDNA by Rapid Amplification of cDNA Ends, based on tryptic and chymotryptic peptide sequences. Partly purified TmXET6.3, expressed in Pichia occurred in N-glycosylated and unglycosylated forms. The quantification of hetero-transglycosylation activities of TmXET6.3 revealed that (1,3,1,4)-, (1,6)- and (1,4)-β-d-glucooligosaccharides were the preferred acceptor substrates, while (1,4)-β-d-xylooligosaccharides, and arabinoxylo- and glucomanno-oligosaccharides were less preferred. The 3D model of TmXET6.3, and bioinformatics analyses of identified and putative plant xyloglucan endotransglycosylases (XETs)/hydrolases (XEHs) of the GH16 family revealed that H94, A104, Q108, K234 and K237 were the key residues that underpinned the acceptor substrate specificity of TmXET6.3. Compared to the wild-type enzyme, the single Q108R and K237T, and double-K234T/K237T and triple-H94Q/A104D/Q108R variants exhibited enhanced hetero-transglycosylation activities with xyloglucan and (1,4)-β-d-glucooligosaccharides, while those with (1,3,1,4)- and (1,6)-β-d-glucooligosaccharides were suppressed, the incorporation of xyloglucan to (1,4)-β-d-glucooligosaccharides by the H94Q variant was influenced most extensively. Structural and biochemical data of non-specific TmXET6.3 presented here extend the classic XET reaction mechanism by which these enzymes operate in plant cell walls. The evaluations of TmXET6.3 transglycosylation activities and the incidence of investigated residues in other members of the GH16 family suggest that a broad acceptor substrate specificity in plant XET enzymes could be more widespread than previously anticipated. Pracoviště Ústav analytické chemie Kontakt Iveta Drobníková, drobnikova@iach.cz, Tel.: 532 290 234 Rok sběru 2020 Elektronická adresa http://hdl.handle.net/11104/0295980
Počet záznamů: 1