Cellobiohydrolase 58 (P.c. Cel 7D) is complementary to the homologous CBHI (T.r. Cel 7A) in enantioseparations
2000 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 898, no 1, 63-74 p.Article in journal (Refereed) Published
Cellobiohydrolase 58 (EC 184.108.40.206, pc. Cel 7D) from Phanerochaete chrysosporium was immobilized on silica and the resulting material, CBH 58-silica, was then used as a chiral stationary phase (CSP) in liquid chromatographic separations of enantiomers. The enantioselectivities obtained on CBH 58-silica were compared with those on CBH I-silica (a phase based on a corresponding cellulase from Trichoderma reesei). CBH 58-silica displayed higher selectivity than CBH I-silica for the more hydrophilic compounds, such as atenolol and metoprolol, although great similarities in chiral separation of beta -adrenergic antagonists were found between the two phases. None of the acidic compounds tested could be resolved on the CBH 58 phase. Moreover, the solutes were retained more on the CBH 58 phase in general, indicating an improved application potential in bioanalysis. Addition of cellobiose or lactose, both of which are inhibitors of cellulases, To the mobile phase impaired the enantioselectivity, indicating an overlap of the enantioselective and catalytic sites. The chiral analytes also functioned as competitive inhibitors and their inhibition constants were determined.
Place, publisher, year, edition, pages
2000. Vol. 898, no 1, 63-74 p.
CHIRAL STATIONARY PHASE, IMMOBILIZED CELLULASE, PHANEROCHAETE-CHRYSOSPORIUM, TRICHODERMA-REESEI, ENANTIOSELECTIVE RETENTION, CAPILLARY ELECTROPHORESIS, BETA-BLOCKERS, I-SILICA, SEPARATION, PROTEIN
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject Biochemistry
IdentifiersURN: urn:nbn:se:uu:diva-87300DOI: 10.1016/S0021-9673(00)00807-4OAI: oai:DiVA.org:uu-87300DiVA: diva2:350