Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 509977
Title A more polar N-terminal helix releases MBP-tagged Thermus thermophilus proline dehydrogenase from tetramer-polymer self-association
Author(s) Huijbers, Mieke M.E.; Berkel, Willem J.H. van
Source Journal of Molecular Catalysis. B, Enzymatic 134 (2016). - ISSN 1381-1177 - p. 340 - 346.
DOI https://doi.org/10.1016/j.molcatb.2016.09.014
Department(s) Biochemistry
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2016
Keyword(s) Flavoprotein - Molecular self-association - Proline dehydrogenase - Protein oligomerization - Thermus thermophilus
Abstract

Proline dehydrogenase (ProDH) is a ubiquitous flavoenzyme involved in the biosynthesis of . l-glutamate. ProDH is of interest for biocatalysis because the protein might be applied in multi-enzyme reactions for the synthesis of structurally complex molecules. We recently demonstrated that the thermotolerant ProDH from . Thermus thermophilus (TtProDH) is overproduced in . Escherichia coli when using maltose-binding protein (MBP) as a solubility tag. However, MBP-TtProDH and MBP-clipped TtProDH are prone to aggregation through non-native self-association. Here we provide evidence that the hydrophobic N-terminal helix of TtProDH is responsible for the self-association process. The more polar MBP-tagged F10E/L12E variant exclusively forms tetramers and exhibits excellent catalytic features over a wide range of temperatures. Understanding the hydrodynamic and catalytic properties of thermostable enzymes is important for the development of industrial biocatalysts as well as for pharmaceutical applications.

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