View source for 6nvx


==Crystal structure of penicillin G acylase from Bacillus sp. FJAT-27231==
<StructureSection load='6nvx' size='340' side='right'caption='[[6nvx]], [[Resolution|resolution]] 1.36&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[6nvx]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Bacillus_sp._FJAT-27231 Bacillus sp. FJAT-27231]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6NVX OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6NVX 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]] 1.36&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=EPE:4-(2-HYDROXYETHYL)-1-PIPERAZINE+ETHANESULFONIC+ACID'>EPE</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</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=6nvx FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6nvx OCA], [https://pdbe.org/6nvx PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6nvx RCSB], [https://www.ebi.ac.uk/pdbsum/6nvx PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6nvx ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/A0A0K9H482_9BACI A0A0K9H482_9BACI] 
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Penicillin G acylase (PGA) catalyzes the hydrolysis of penicillin G to 6-aminopenicillanic acid and phenylacetic acid, which provides the precursor for most semisynthetic penicillins. Most applications rely on PGAs from Gram-negative bacteria. Here we describe the first three crystal structures for PGAs from Gram-positive Bacilli and their utilization in protein engineering experiments for the manipulation of their thermostability. PGAs from Bacillus megaterium (BmPGA, Tm = 56.0 degrees C), Bacillus thermotolerans (BtPGA, Tm = 64.5 degrees C), and Bacillus sp. FJAT-27231 (FJAT-PGA, Tm = 74.3 degrees C) were recombinantly produced with B. megaterium, secreted, purified to apparent heterogeneity, and crystallized. Structures with resolutions of 2.20 A (BmPGA), 2.27 A (BtPGA), and 1.36 A (FJAT-PGA) were obtained. They revealed high overall similarity, reflecting the high identity of up to approx. 75%. Notably, the active center displays a deletion of more than ten residues with respect to PGAs from Gram-negatives. This enlarges the substrate binding site and may indicate a different substrate spectrum. Based on the structures, ten single-chain FJAT-PGAs carrying artificial linkers were produced. However, in all cases, complete linker cleavage was observed. While thermostability remained in the wild-type range, the enzymatic activity dropped between 30 and 60%. Furthermore, four hybrid PGAs carrying subunits from two different enzymes were successfully produced. Their thermostabilities mostly lay between the values of the two mother enzymes. For one PGA increased, enzyme activity was observed. Overall, the three novel PGA structures combined with initial protein engineering experiments provide the basis for establishment of new PGA-based biotechnological processes.

Crystal structures and protein engineering of three different penicillin G acylases from Gram-positive bacteria with different thermostability.,Mayer J, Pippel J, Gunther G, Muller C, Lauermann A, Knuuti T, Blankenfeldt W, Jahn D, Biedendieck R Appl Microbiol Biotechnol. 2019 Jun 21. pii: 10.1007/s00253-019-09977-8. doi:, 10.1007/s00253-019-09977-8. PMID:31227867<ref>PMID:31227867</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 6nvx" style="background-color:#fffaf0;"></div>

==See Also==
*[[Penicillin acylase|Penicillin acylase]]
== References ==
<references/>
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</StructureSection>
[[Category: Bacillus sp. FJAT-27231]]
[[Category: Large Structures]]
[[Category: Blankenfeldt W]]