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Design of α-Keto Carboxylic Acid Dimers by Domain Recombination of Nonribosomal Peptide Synthetase (NRPS)-Like Enzymes.
Org Lett
January 2019
Institut für Pharmazeutische Biologie und Biotechnologie , Philipps-Universität Marburg, Robert-Koch Straße 4 , Marburg 35037 , Germany.
Nonribosomal peptide synthetase (NRPS)-like enzymes comprising A-T-TE architectures catalyze the dimerization of α-keto carboxylic acids leading to the formation of hydroxybenzoquinones or lactones. Domain change experiments with five enzymes revealed that A and A-T domains of phenyl or 4-hydroxyphenyl pyruvate-using enzymes can be effectively used by the TE domains of other enzymes. Even the A and A-T domains of an indolyl hydroxybenzoquinone synthase were successfully recombined with TE domains of a phenyl and a 4-hydroxyphenyl pyruvate-activating enzyme.
View Article and Find Full Text PDFProduction of α-keto carboxylic acid dimers in yeast by overexpression of NRPS-like genes from Aspergillus terreus.
Appl Microbiol Biotechnol
February 2018
Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Robert-Koch-Straße 4, 35037, Marburg, Germany.
Non-ribosomal peptide synthetases (NRPSs) are key enzymes in microorganisms for the assembly of peptide backbones of biologically and pharmacologically active natural products. The monomodular NRPS-like enzymes comprise often an adenylation (A), a thiolation (T), and a thioesterase (TE) domain. In contrast to the NRPSs, they do not contain any condensation domain and usually catalyze a dimerization of α-keto carboxylic acids and thereby provide diverse scaffolds for further modifications.
View Article and Find Full Text PDFTwo-dimensional capillary zone electrophoresis-mass spectrometry for the characterization of intact monoclonal antibody charge variants, including deamidation products.
Anal Bioanal Chem
October 2017
Faculty of Chemistry, Aalen University, Beethovenstr. 1, 73430, Aalen, Germany.
Capillary zone electrophoresis (CZE) is a powerful tool that is progressively being applied for the separation of monoclonal antibody (mAb) charge variants. Mass spectrometry (MS) is the desired detection method concerning identification of mAb variants. In biopharmaceutical applications, there exist optimized and validated electrolyte systems for mAb variant quantification.
View Article and Find Full Text PDFInterleukin-4 Protects Dopaminergic Neurons but Is Dispensable for MPTP-Induced Neurodegeneration .
Front Mol Neurosci
March 2017
Department of Molecular Embryology, Institute for Anatomy and Cell Biology, Faculty of Medicine, University of Freiburg Freiburg, Germany.
Microglia are involved in physiological as well as neuropathological processes in the central nervous system (CNS). Their functional states are often referred to as M1-like and M2-like activation, and are believed to contribute to neuroinflammation-mediated neurodegeneration or neuroprotection, respectively. Parkinson's disease (PD) is one the most common neurodegenerative disease and is characterized by the progressive loss of midbrain dopaminergic (mDA) neurons in the substantia nigra resulting in bradykinesia, tremor, and rigidity.
View Article and Find Full Text PDFInterference-free mass spectrometric detection of capillary isoelectric focused proteins, including charge variants of a model monoclonal antibody.
Electrophoresis
March 2017
Faculty of Chemistry, Aalen University, Aalen, Germany.
CIEF represents an elegant technique especially for the separation of structural similar analytes, whereas MS is a state-of-the-art instrumentation for the identification and characterization of biomolecules. The combination of both techniques can be realized by hyphenating CIEF with CZE-ESI-MS applying a mechanical valve. During the CZE step, the remaining ESI-interfering components of the CIEF electrolyte are separated from the analytes prior to MS detection.
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