Kinetic analyses of retaining endo-(Xylo)glucanases from plant and microbial sources using new chromogenic xylogluco-oligosaccharide aryl glycosides. Ibatullin, F., Baumann, M., Greffe, L., & Brumer, H. Biochemistry, 47(29):7762-7769, 2008. cited By 24
Paper doi abstract bibtex A library of phenyl β-glycosides of xylogluco-oligosaccharides was synthesized via a chemoenzymatic approach to produce new, specific substrates for xyloglucanases. Tamarind xyloglucan was completely hydrolyzed to four, variably galactosylated component oligosaccharides based on Glc4 backbones, using a Trichoderma endo-glucanase mixture. Oligosaccharide complexity could be further reduced by β-galactosidase treament. Subsequent per-O-acetylation, α-bromination, phase-transfer glycosylation, and Zemplén deprotection yielded phenyl glycosides of XXXG and XLLG oligosaccharides with a broad range of aglycon pKa values. Kinetic and product analysis of the action of the archetypal plant endo-xyloglucanase, Tropaeolum majus NXG1, on these compounds indicated that formation of the glycosyl-enzyme intermediate was rate-limiting in the case of phenol leaving groups with pKa values of >7, leading exclusively to substrate hydrolysis. Conversely, substrates with aglycon pKa values of 5.4 gave rise to a significant amount of transglycosylation products, indicating a change in the relative rates of formation and breakdown of the glycosyl-enzyme intermediate for these faster substrates. Notably, comparison of the initial rates of XXXG-Ar and XLLG-Ar conversion indicated that catalysis by TmNXG1 was essentially insensitive to the presence of galactose in the negative subsites for all leaving groups. More broadly, analysis of a selection of enzymes from CAZy families GH 5, 12, and 16 indicated that the phenyl glycosides are substrates for anomeric configuration-retaining endo-xyloglucanases but are not substrates for strict xyloglucan endo-transglycosylases (XETs). The relative activities of the GH 5, 12, and 16 endo-xyloglucanases toward GGGG-CNP, XXXG-CNP, and XLLG-CNP reflected those observed using analogous high molar mass polysaccharides. These new chromogenic substrates may thus find wide application in the discovery, screening, and detailed kinetic analysis of new xyloglucan-active enzymes. © 2008 American Chemical Society.
@ARTICLE{Ibatullin20087762,
author={Ibatullin, F.M. and Baumann, M.J. and Greffe, L. and Brumer, H.},
title={Kinetic analyses of retaining endo-(Xylo)glucanases from plant and microbial sources using new chromogenic xylogluco-oligosaccharide aryl glycosides},
journal={Biochemistry},
year={2008},
volume={47},
number={29},
pages={7762-7769},
doi={10.1021/bi8009168},
note={cited By 24},
url={https://www.scopus.com/inward/record.uri?eid=2-s2.0-47649125408&doi=10.1021%2fbi8009168&partnerID=40&md5=66aea507deaaf913166a027f9f76526d},
affiliation={School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Centre, SE-106 91 Stockholm, Sweden; Petersburg Nuclear Physics Institute, Molecular and Radiation Biology Division, Russian Academy of Science, Gatchina, St. Petersburg 188300, Russian Federation},
abstract={A library of phenyl β-glycosides of xylogluco-oligosaccharides was synthesized via a chemoenzymatic approach to produce new, specific substrates for xyloglucanases. Tamarind xyloglucan was completely hydrolyzed to four, variably galactosylated component oligosaccharides based on Glc4 backbones, using a Trichoderma endo-glucanase mixture. Oligosaccharide complexity could be further reduced by β-galactosidase treament. Subsequent per-O-acetylation, α-bromination, phase-transfer glycosylation, and Zemplén deprotection yielded phenyl glycosides of XXXG and XLLG oligosaccharides with a broad range of aglycon pKa values. Kinetic and product analysis of the action of the archetypal plant endo-xyloglucanase, Tropaeolum majus NXG1, on these compounds indicated that formation of the glycosyl-enzyme intermediate was rate-limiting in the case of phenol leaving groups with pKa values of >7, leading exclusively to substrate hydrolysis. Conversely, substrates with aglycon pKa values of 5.4 gave rise to a significant amount of transglycosylation products, indicating a change in the relative rates of formation and breakdown of the glycosyl-enzyme intermediate for these faster substrates. Notably, comparison of the initial rates of XXXG-Ar and XLLG-Ar conversion indicated that catalysis by TmNXG1 was essentially insensitive to the presence of galactose in the negative subsites for all leaving groups. More broadly, analysis of a selection of enzymes from CAZy families GH 5, 12, and 16 indicated that the phenyl glycosides are substrates for anomeric configuration-retaining endo-xyloglucanases but are not substrates for strict xyloglucan endo-transglycosylases (XETs). The relative activities of the GH 5, 12, and 16 endo-xyloglucanases toward GGGG-CNP, XXXG-CNP, and XLLG-CNP reflected those observed using analogous high molar mass polysaccharides. These new chromogenic substrates may thus find wide application in the discovery, screening, and detailed kinetic analysis of new xyloglucan-active enzymes. © 2008 American Chemical Society.},
correspondence_address1={Brumer, H.; School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Centre, SE-106 91 Stockholm, Sweden; email: harry@biotech.kth.se},
issn={00062960},
coden={BICHA},
pubmed_id={18627132},
language={English},
abbrev_source_title={Biochemistry},
document_type={Article},
source={Scopus},
}
Downloads: 0
{"_id":"guBEz9BCykTDDvmrk","bibbaseid":"ibatullin-baumann-greffe-brumer-kineticanalysesofretainingendoxyloglucanasesfromplantandmicrobialsourcesusingnewchromogenicxyloglucooligosaccharidearylglycosides-2008","author_short":["Ibatullin, F.","Baumann, M.","Greffe, L.","Brumer, H."],"bibdata":{"bibtype":"article","type":"article","author":[{"propositions":[],"lastnames":["Ibatullin"],"firstnames":["F.M."],"suffixes":[]},{"propositions":[],"lastnames":["Baumann"],"firstnames":["M.J."],"suffixes":[]},{"propositions":[],"lastnames":["Greffe"],"firstnames":["L."],"suffixes":[]},{"propositions":[],"lastnames":["Brumer"],"firstnames":["H."],"suffixes":[]}],"title":"Kinetic analyses of retaining endo-(Xylo)glucanases from plant and microbial sources using new chromogenic xylogluco-oligosaccharide aryl glycosides","journal":"Biochemistry","year":"2008","volume":"47","number":"29","pages":"7762-7769","doi":"10.1021/bi8009168","note":"cited By 24","url":"https://www.scopus.com/inward/record.uri?eid=2-s2.0-47649125408&doi=10.1021%2fbi8009168&partnerID=40&md5=66aea507deaaf913166a027f9f76526d","affiliation":"School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Centre, SE-106 91 Stockholm, Sweden; Petersburg Nuclear Physics Institute, Molecular and Radiation Biology Division, Russian Academy of Science, Gatchina, St. Petersburg 188300, Russian Federation","abstract":"A library of phenyl β-glycosides of xylogluco-oligosaccharides was synthesized via a chemoenzymatic approach to produce new, specific substrates for xyloglucanases. Tamarind xyloglucan was completely hydrolyzed to four, variably galactosylated component oligosaccharides based on Glc4 backbones, using a Trichoderma endo-glucanase mixture. Oligosaccharide complexity could be further reduced by β-galactosidase treament. Subsequent per-O-acetylation, α-bromination, phase-transfer glycosylation, and Zemplén deprotection yielded phenyl glycosides of XXXG and XLLG oligosaccharides with a broad range of aglycon pKa values. Kinetic and product analysis of the action of the archetypal plant endo-xyloglucanase, Tropaeolum majus NXG1, on these compounds indicated that formation of the glycosyl-enzyme intermediate was rate-limiting in the case of phenol leaving groups with pKa values of >7, leading exclusively to substrate hydrolysis. Conversely, substrates with aglycon pKa values of 5.4 gave rise to a significant amount of transglycosylation products, indicating a change in the relative rates of formation and breakdown of the glycosyl-enzyme intermediate for these faster substrates. Notably, comparison of the initial rates of XXXG-Ar and XLLG-Ar conversion indicated that catalysis by TmNXG1 was essentially insensitive to the presence of galactose in the negative subsites for all leaving groups. More broadly, analysis of a selection of enzymes from CAZy families GH 5, 12, and 16 indicated that the phenyl glycosides are substrates for anomeric configuration-retaining endo-xyloglucanases but are not substrates for strict xyloglucan endo-transglycosylases (XETs). The relative activities of the GH 5, 12, and 16 endo-xyloglucanases toward GGGG-CNP, XXXG-CNP, and XLLG-CNP reflected those observed using analogous high molar mass polysaccharides. These new chromogenic substrates may thus find wide application in the discovery, screening, and detailed kinetic analysis of new xyloglucan-active enzymes. © 2008 American Chemical Society.","correspondence_address1":"Brumer, H.; School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Centre, SE-106 91 Stockholm, Sweden; email: harry@biotech.kth.se","issn":"00062960","coden":"BICHA","pubmed_id":"18627132","language":"English","abbrev_source_title":"Biochemistry","document_type":"Article","source":"Scopus","bibtex":"@ARTICLE{Ibatullin20087762,\r\nauthor={Ibatullin, F.M. and Baumann, M.J. and Greffe, L. and Brumer, H.},\r\ntitle={Kinetic analyses of retaining endo-(Xylo)glucanases from plant and microbial sources using new chromogenic xylogluco-oligosaccharide aryl glycosides},\r\njournal={Biochemistry},\r\nyear={2008},\r\nvolume={47},\r\nnumber={29},\r\npages={7762-7769},\r\ndoi={10.1021/bi8009168},\r\nnote={cited By 24},\r\nurl={https://www.scopus.com/inward/record.uri?eid=2-s2.0-47649125408&doi=10.1021%2fbi8009168&partnerID=40&md5=66aea507deaaf913166a027f9f76526d},\r\naffiliation={School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Centre, SE-106 91 Stockholm, Sweden; Petersburg Nuclear Physics Institute, Molecular and Radiation Biology Division, Russian Academy of Science, Gatchina, St. Petersburg 188300, Russian Federation},\r\nabstract={A library of phenyl β-glycosides of xylogluco-oligosaccharides was synthesized via a chemoenzymatic approach to produce new, specific substrates for xyloglucanases. Tamarind xyloglucan was completely hydrolyzed to four, variably galactosylated component oligosaccharides based on Glc4 backbones, using a Trichoderma endo-glucanase mixture. Oligosaccharide complexity could be further reduced by β-galactosidase treament. Subsequent per-O-acetylation, α-bromination, phase-transfer glycosylation, and Zemplén deprotection yielded phenyl glycosides of XXXG and XLLG oligosaccharides with a broad range of aglycon pKa values. Kinetic and product analysis of the action of the archetypal plant endo-xyloglucanase, Tropaeolum majus NXG1, on these compounds indicated that formation of the glycosyl-enzyme intermediate was rate-limiting in the case of phenol leaving groups with pKa values of >7, leading exclusively to substrate hydrolysis. Conversely, substrates with aglycon pKa values of 5.4 gave rise to a significant amount of transglycosylation products, indicating a change in the relative rates of formation and breakdown of the glycosyl-enzyme intermediate for these faster substrates. Notably, comparison of the initial rates of XXXG-Ar and XLLG-Ar conversion indicated that catalysis by TmNXG1 was essentially insensitive to the presence of galactose in the negative subsites for all leaving groups. More broadly, analysis of a selection of enzymes from CAZy families GH 5, 12, and 16 indicated that the phenyl glycosides are substrates for anomeric configuration-retaining endo-xyloglucanases but are not substrates for strict xyloglucan endo-transglycosylases (XETs). The relative activities of the GH 5, 12, and 16 endo-xyloglucanases toward GGGG-CNP, XXXG-CNP, and XLLG-CNP reflected those observed using analogous high molar mass polysaccharides. These new chromogenic substrates may thus find wide application in the discovery, screening, and detailed kinetic analysis of new xyloglucan-active enzymes. © 2008 American Chemical Society.},\r\ncorrespondence_address1={Brumer, H.; School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Centre, SE-106 91 Stockholm, Sweden; email: harry@biotech.kth.se},\r\nissn={00062960},\r\ncoden={BICHA},\r\npubmed_id={18627132},\r\nlanguage={English},\r\nabbrev_source_title={Biochemistry},\r\ndocument_type={Article},\r\nsource={Scopus},\r\n}\r\n\r\n","author_short":["Ibatullin, F.","Baumann, M.","Greffe, L.","Brumer, H."],"key":"Ibatullin20087762","id":"Ibatullin20087762","bibbaseid":"ibatullin-baumann-greffe-brumer-kineticanalysesofretainingendoxyloglucanasesfromplantandmicrobialsourcesusingnewchromogenicxyloglucooligosaccharidearylglycosides-2008","role":"author","urls":{"Paper":"https://www.scopus.com/inward/record.uri?eid=2-s2.0-47649125408&doi=10.1021%2fbi8009168&partnerID=40&md5=66aea507deaaf913166a027f9f76526d"},"metadata":{"authorlinks":{}}},"bibtype":"article","biburl":"https://bio.pnpi.nrcki.ru/wp-content/uploads/2019/12/lbmx_2019_10.txt","dataSources":["5udpPf7ooeTjMNNMd"],"keywords":[],"search_terms":["kinetic","analyses","retaining","endo","xylo","glucanases","plant","microbial","sources","using","new","chromogenic","xylogluco","oligosaccharide","aryl","glycosides","ibatullin","baumann","greffe","brumer"],"title":"Kinetic analyses of retaining endo-(Xylo)glucanases from plant and microbial sources using new chromogenic xylogluco-oligosaccharide aryl glycosides","year":2008}