BioChem - DOS

Solution Structure of a Bacterial Microcompartment Targeting Peptide and Its Application in the Construction of an Ethanol Bioreactor

ACS Synthetic biology

Lawrence, Andrew D.; Frank, Stefanie; Newnham, Sarah; Lee, Matthew J.; Brown, Ian R.; Xue, Wei-Feng; Rowe, Michelle L.; Mulvihill, Daniel P.; Prentice, Michael B.; Howard, Mark J.; Warren, Martin J.

<P>Targeting of proteins to bacterial microcompartments (BMCs) is mediated by an 18-amino-acid peptide sequence. Herein, we report the solution structure of the N-terminal targeting peptide (P18) of PduP, the aldehyde dehydrogenase associated with the 1,2-propanediol utilization metabolosome from <I>Citrobacter freundii</I>. The solution structure reveals the peptide to have a well-defined helical conformation along its whole length. Saturation transfer difference and transferred NOE NMR has highlighted the observed interaction surface on the peptide with its main interacting shell protein, PduK. By tagging both a pyruvate decarboxylase and an alcohol dehydrogenase with targeting peptides, it has been possible to direct these enzymes to empty BMCs <I>in vivo</I> and to generate an ethanol bioreactor. Not only are the purified, redesigned BMCs able to transform pyruvate into ethanol efficiently, but the strains containing the modified BMCs produce elevated levels of alcohol.</P><P><B>Graphic Abstract</B><BR><IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/asbcd6/2014/asbcd6.2014.3.issue-7/sb4001118/production/images/medium/sb-2013-001118_0010.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/sb4001118'>ACS Electronic Supporting Info</A></P>

2014

American Chemical Society

2161-5063

3

7

pp.454-465

10.1021/sb4001118

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART70019002

https://europepmc.org/articles/pmc4880047?pdf=render

metabolic engineering; compartmentalization; propanediol utilization; synthetic biology