View source for 7exs


==Thermomicrobium roseum sarcosine oxidase mutant - S320R==
<StructureSection load='7exs' size='340' side='right'caption='[[7exs]], [[Resolution|resolution]] 1.42&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[7exs]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Thermomicrobium_roseum_DSM_5159 Thermomicrobium roseum DSM 5159]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7EXS OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7EXS FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.42&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene></td></tr>
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=7exs FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7exs OCA], [https://pdbe.org/7exs PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7exs RCSB], [https://www.ebi.ac.uk/pdbsum/7exs PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7exs ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/B9L163_THERP B9L163_THERP] 
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
N-demethylases have been reported to remove the methyl groups on primary or secondary amines, which could further affect the properties and functions of biomacromolecules or chemical compounds; however, the substrate scope and the robustness of N-demethylases have not been systematically investigated. Here we report the recreation of natural evolution in key microdomains of the Thermomicrobium roseum sarcosine oxidase (TrSOX), an N-demethylase with marked stability (melting temperature over 100 degrees C) and enantioselectivity, for enhanced substrate scope and catalytic efficiency on -C-N- bonds. We obtained the structure of TrSOX by crystallization and X-ray diffraction (XRD) for the initial framework. The natural evolution in the non-conserved residues of key microdomains-including the catalytic loop, coenzyme pocket, substrate pocket, and entrance site-were then identified using ancestral sequence reconstruction (ASR), and the substitutions that accrued during natural evolution were recreated by site-directed mutagenesis. The single and double substitution variants catalyzed the N-demethylation of N-methyl-L-amino acids up to 1800- and 6000-fold faster than the wild type, respectively. Additionally, these single substitution variants catalyzed the terminal N-demethylation of non-amino acid compounds and the oxidation of the main chain -C-N- bond to a -C=N- bond in the nitrogen-containing heterocycle. Notably, these variants retained the enantioselectivity and stability of the initial framework. We conclude that the variants of TrSOX are of great potential use in N-methyl enantiomer resolution, main-chain Schiff base synthesis, and alkaloid modification or degradation.

Recreating the natural evolutionary trend in key microdomains provides an effective strategy for engineering of a thermomicrobial N-demethylase.,Xin Y, Shen C, Tang M, Guo Z, Shi Y, Gu Z, Shao J, Zhang L J Biol Chem. 2022 Feb 3:101656. doi: 10.1016/j.jbc.2022.101656. PMID:35124004<ref>PMID:35124004</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 7exs" style="background-color:#fffaf0;"></div>

==See Also==
*[[Sarcosine oxidase|Sarcosine oxidase]]
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Large Structures]]
[[Category: Thermomicrobium roseum DSM 5159]]
[[Category: Gu ZH]]
[[Category: Guo ZT]]
[[Category: Shao J]]
[[Category: Shen C]]
[[Category: Shi Y]]
[[Category: Tang MW]]
[[Category: Xin Y]]
[[Category: Zhang L]]