Biochemical properties of GH94 cellodextrin phosphorylase THA_1941 from a thermophilic eubacterium Thermosipho africanus TCF52B with cellobiose phosphorylase activity

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dc.contributor.authorWu, Yuanyuanen
dc.contributor.authorMao, Guotaoen
dc.contributor.authorFan, Haiyanen
dc.contributor.authorSong, Andongen
dc.contributor.authorZhang, Yi-Heng Percivalen
dc.contributor.authorChen, Honggeen
dc.contributor.departmentBiological Systems Engineeringen
dc.date.accessioned2019-01-08T15:53:29Zen
dc.date.available2019-01-08T15:53:29Zen
dc.date.issued2017-07-07en
dc.description.abstractA hypothetic gene (THA_1941) encoding a putative cellobiose phosphorylase (CBP) from Thermosipho africanus TCF52B has very low amino acid identities (less than 12%) to all known GH94 enzymes. This gene was cloned and over-expressed in Escherichia coli BL21(DE3). The recombinant protein was hypothesized to be a CBP enzyme and it showed an optimum temperature of 75 degrees C and an optimum pH of 7.5. Beyond its CBP activity, this enzyme can use cellobiose and long-chain cellodextrins with a degree of polymerization of greater than two as a glucose acceptor, releasing phosphate from glucose 1-phosphate. The catalytic efficiencies (k(cat)/K-m) indicated that cellotetraose and cellopentaose were the best substrates for the phosphorolytic and reverse synthetic reactions, respectively. These results suggested that this enzyme was the first enzyme having both cellodextrin and cellobiose phosphorylases activities. Because it preferred cellobiose and cellodextrins to glucose in the synthetic direction, it was categorized as a cellodextrin phosphorylase (CDP). Due to its unique ability of the reverse synthetic reaction, this enzyme could be a potential catalyst for the synthesis of various oligosaccharides. The speculative function of this CDP in the carbohydrate metabolism of T. africanus TCF52B was also discussed.en
dc.description.notesThe authors thank Dr. Chun You in Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences for his help in the verification of enzyme properties. This work was supported by the National Natural Science Foundation of China (No. 31571775) to H.C.en
dc.description.sponsorshipNational Natural Science Foundation of China [31571775]en
dc.format.extent12en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1038/s41598-017-05289-xen
dc.identifier.issn2045-2322en
dc.identifier.other4849en
dc.identifier.pmid28687766en
dc.identifier.urihttp://hdl.handle.net/10919/86634en
dc.identifier.volume7en
dc.language.isoenen
dc.publisherSpringer Natureen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectclostridium-thermocellumen
dc.subjectruminococcus-albusen
dc.subjectoligosaccharide synthesisen
dc.subjectreverse phosphorolysisen
dc.subjectescherichia-colien
dc.subjectcellulomonas-udaen
dc.subjectpurificationen
dc.subjectmechanismen
dc.subjectcleavageen
dc.subjectfermentationen
dc.titleBiochemical properties of GH94 cellodextrin phosphorylase THA_1941 from a thermophilic eubacterium Thermosipho africanus TCF52B with cellobiose phosphorylase activityen
dc.title.serialScientific Reportsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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