caa a22 4500 445385103 CHVBK 20180317143030.0 cr unu---uuuuu 170323e20111201xx s 000 0 eng 10.1007/s10924-011-0345-x doi (NATIONALLICENCE)springer-10.1007/s10924-011-0345-x A Novel Affinity Chromatographic Material for the Purification of Extracellular Polyhydroxybutyrate Depolymerases [Elektronische Daten] [Christos Papaneophytou, Anastasia Pantazaki] A novel affinity chromatographic material, which is composed of silica matrix, coated with polyhydroxybutyrate (PHB) powder, suitable for the purification of PHB depolymerases, was developed. The surface morphology of the PHB-silica coated particles (silica-PHB composite particles) was examined by scanning electron microscopy and revealed a successful uniform coating of silica particles with PHB. Moreover, the complex of these materials retained its homogeneity even after incubation at 80°C for 6h, whereas the strong binding of PHB on silica surface was further verified by thermal gravimetric analysis and by PHB extraction- from silica surface- experiments. This novel material was demonstrated to be suitable for both, the one-step on-batch and on-column purification of Thermus thermophilus extracellular PHB depolymerase. The enzyme exhibited higher affinity against the composite of silica-PHB particles than PHB powder, since the one-step purification-fold and the overall recovery of the enzyme were 2.8 and 4 times higher respectively, in the first case. Reusability of the silica-PHB composites particles was examined by determining the recoveries of PHB depolymerase. The enzyme recoveries were ranged from 30 to 35% for the first five uses, whereas for further uses recoveries gradually dropped to 15-18% indicating that the particles could be used repeatedly for five times. This material could be also a suitable support for lipases or other proteins that exhibit strong affinity to hydrophobic materials. Springer Science+Business Media, LLC, 2011 Affinity chromatography nationallicence Silica-PHB composite particles nationallicence Extracellular PHB-depolymerase nationallicence Thermus thermophilus HB8 nationallicence Papaneophytou Christos Department of Chemistry, Laboratory of Biochemistry, Aristotle University of Thessaloniki, 54124, Thessaloniki, Greece aut Pantazaki Anastasia Department of Chemistry, Laboratory of Biochemistry, Aristotle University of Thessaloniki, 54124, Thessaloniki, Greece aut Journal of Polymers and the Environment Springer US; http://www.springer-ny.com 19/4(2011-12-01), 876-886 1566-2543 19:4<876 2011 19 10924 https://doi.org/10.1007/s10924-011-0345-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10924-011-0345-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Papaneophytou Christos Department of Chemistry, Laboratory of Biochemistry, Aristotle University of Thessaloniki, 54124, Thessaloniki, Greece aut NATIONALLICENCE 700 1- Pantazaki Anastasia Department of Chemistry, Laboratory of Biochemistry, Aristotle University of Thessaloniki, 54124, Thessaloniki, Greece aut NATIONALLICENCE 773 0- Journal of Polymers and the Environment Springer US; http://www.springer-ny.com 19/4(2011-12-01), 876-886 1566-2543 19:4<876 2011 19 10924 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer