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      Activation of Recombinantly Expressed l-Amino Acid Oxidase from Rhizoctonia solani by Sodium Dodecyl Sulfate

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          Abstract

          l-Amino acid oxidases ( l-AAO) catalyze the oxidative deamination of l-amino acids to the corresponding α-keto acids. The non-covalently bound cofactor FAD is reoxidized by oxygen under formation of hydrogen peroxide. We expressed an active l-AAO from the fungus Rhizoctonia solani as a fusion protein in E. coli. Treatment with small amounts of the detergent sodium dodecyl sulfate (SDS) stimulated the activity of the enzyme strongly. Here, we investigated whether other detergents and amphiphilic molecules activate 9His- rsLAAO1. We found that 9His- rsLAAO1 was also activated by sodium tetradecyl sulfate. Other detergents and fatty acids were not effective. Moreover, effects of SDS on the oligomerization state and the protein structure were analyzed. Native and SDS-activated 9His- rsLAAO1 behaved as dimers by size-exclusion chromatography. SDS treatment induced an increase in hydrodynamic radius as observed by size-exclusion chromatography and dynamic light scattering. The activated enzyme showed accelerated thermal inactivation and an exposure of additional protease sites. Changes in tryptophan fluorescence point to a more hydrophilic environment. Moreover, FAD fluorescence increased and a lower concentration of sulfites was sufficient to form adducts with FAD. Taken together, these data point towards a more open conformation of SDS-activated l-amino acid oxidase facilitating access to the active site.

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          Analysis of Macromolecular Polydispersity in Intensity Correlation Spectroscopy: The Method of Cumulants

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            A constrained regularization method for inverting data represented by linear algebraic or integral equations

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              CONTIN: A general purpose constrained regularization program for inverting noisy linear algebraic and integral equations

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                Author and article information

                Journal
                Molecules
                Molecules
                molecules
                Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry
                MDPI
                1420-3049
                20 December 2017
                December 2017
                : 22
                : 12
                : 2272
                Affiliations
                [1 ]Biochemistry III, Department of Chemistry, Universitätsstrasse 25, Bielefeld University, 33615 Bielefeld, Germany; katharina.hahn@ 123456uni-bielefeld.de (K.H.); sbloess@ 123456uni-bielefeld.de (S.B.)
                [2 ]Physical and Biophysical Chemistry, Department of Chemistry, Universitätsstrasse 25, Bielefeld University, 33615 Bielefeld, Germany; yvonne.hertle@ 123456uni-bielefeld.de (Y.H.); tilman.kottke@ 123456uni-bielefeld.de (T.K.); thomas.hellweg@ 123456uni-bielefeld.de (T.H.)
                Author notes
                [* ]Correspondence: gabriele.mollard@ 123456uni-bielefeld.de ; Tel.: +49-521-106-2081
                Author information
                https://orcid.org/0000-0001-8080-9579
                Article
                molecules-22-02272
                10.3390/molecules22122272
                6149798
                29261108
                4118c4e6-4a79-4139-871c-518cffda2e04
                © 2017 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 10 November 2017
                : 18 December 2017
                Categories
                Article

                l-amino acid oxidase,activation,sds,active conformation,conformational change,photon correlation spectroscopy (pcs)

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