Christen E.H.,Albert Ludwigs University of Freiburg |
Christen E.H.,ETH Zurich |
Gubeli R.J.,Albert Ludwigs University of Freiburg |
Kaufmann B.,Albert Ludwigs University of Freiburg |
And 7 more authors.
Organic and Biomolecular Chemistry | Year: 2012
The Cu(i)-catalyzed cycloaddition of terminal azides and alkynes (click chemistry) represents a highly specific reaction for the functionalization of biomolecules with chemical moieties such as dyes or polymer matrices. In this study we evaluate the use of bicinchoninic acid (BCA) as a ligand for Cu(i) under physiological reaction conditions. We demonstrate that the BCA-Cu(i)-complex represents an efficient catalyst for the conjugation of fluorophores or biotin to alkyne- or azide-functionalized proteins resulting in increased or at least equal reaction yields compared to commonly used catalysts like Cu(i) in complex with TBTA (tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl] amine) or BPAA (bathophenanthroline disulfonic acid). The stabilization of Cu(i) with BCA represents a new strategy for achieving highly efficient bioconjugation reactions under physiological conditions in many application fields. © 2012 The Royal Society of Chemistry.
Acevedo-Rocha C.G.,Max-Planck-Institut fur Kohlenforschung |
Acevedo-Rocha C.G.,University of Marburg |
Hoesl M.G.,TU Berlin |
Nehring S.,TU Berlin |
And 5 more authors.
Catalysis Science and Technology | Year: 2013
The incorporation of several non-canonical amino acids into the Thermoanaerobacter thermohydrosulfuricus lipase confers not only activity enhancement upon treatment with organic solvents (by up to 450%) and surfactants (resp. 1630%), but also protective effects against protein reducing (resp. 140%), alkylating (resp. 160%), and denaturing (resp.190%) agents as well as inhibitors (resp. 40%). This approach offers novel chemically diversified biocatalysts for hostile environments. © 2013 The Royal Society of Chemistry.
Fliedl L.,ACIB |
Grillari J.,ACIB |
Grillari J.,University of Natural Resources and Life Sciences, Vienna |
Grillari-Voglauer R.,ACIB |
Grillari-Voglauer R.,University of Natural Resources and Life Sciences, Vienna
New Biotechnology | Year: 2015
The market of recombinant proteins as human pharmaceuticals has surpassed annual revenues of more than 150 billion dollars. The marketed proteins are often complex in terms of post-translational modifications and conventional hosts have shown weaknesses in terms of quality of these recombinant proteins. Especially the non-human glycopatterns leading to immunogenicity or shortened in vivo half-life have gained attention over the past decade. Therefore, production cell lines with better or novel characteristics are required and human cell lines seem to be the most genuine and logical choice. Thus, several human cell lines have been used to generate biopharmaceuticals. We here present an overview of such examples and highlight their promise for biopharmaceutical production processes of the future. © 2014 Elsevier B.V.
Ribitsch D.,ACIB |
Acero E.H.,ACIB |
Greimel K.,ACIB |
Eiteljoerg I.,ACIB |
And 6 more authors.
Biocatalysis and Biotransformation | Year: 2012
A new cutinase from Thermobifida alba (Tha-Cut1) was cloned and characterized for polyethylene terephthalate (PET) hydrolysis. Tha-Cut1 showed a high degree of identity to a T. cellulolysitica cutinase with only four amino acid differences outside the active site area, according to modeling data. Yet, Tha-Cut1 was more active in terms of PET surface hydrolysis leading to considerable improvement in hydrophilicity quantified based on a decrease of the water contact angle from 87.7° to 45.0°. The introduction of new carboxyl groups was confirmed and measured after esterification with the fluorescent reagent alkyl bromide, 2-(bromomethyl) naphthalene (BrNP), resulting in a fluorescence emission intensity increase from 980 to 1420 a.u. On the soluble model substrates p-nitrophenyl acetate (PNPA) and p-nitrophenyl butyrate (PNPB), the cutinase showed Km values of 213 and 1933 μM and kcat values of 2.72 and 6.03 s -1 respectively. The substrate specificity was investigated with bis(benzoyloxyethyl)terephthalate (3PET) and Tha-Cut1 was shown to release primarily 2-hydroxyethyl benzoate. This contrasts with the well-studied Humicula insolens cutinase which preferentially liberates terminal benzoic acid from 3PET. © 2012 Informa UK, Ltd.
Krempl P.M.,ACIB |
Krempl P.M.,University of Graz |
Mairhofer J.,ACIB |
Mairhofer J.,Institute of Applied Microbiology |
And 4 more authors.
Bioinformatics | Year: 2012
We present a plug-in for Pathway Tools, an integrated systems biology software to create, maintain and query Pathway/Genome Databases. Fully integrated into the graphical user interface and menu, this plug-in extends the application's functionality by the ability to create multiple sequence alignments, systematically annotate insertion sequence (IS) elements and analyse their activity by cross-species comparison tools. Microarray probes can be automatically mapped to target genes, and expression data obtained with these arrays can be transformed into input formats needed to visualize them in the various omics viewers of Pathway Tools. The plug-in API itself allows developers to integrate their own functions into the Pathway Tools menu. © The Author 2012. Published by Oxford University Press. All rights reserved.