BioChem - DOS

High-throughput screen for sorting cells capable of producing the biofuel feedstock botryococcene

Organic & biomolecular chemistry

Chacko, Allen R.; Amster, Daniel E.; Johnson, Tyler E.; Newman, Sara R.; Gladchuk, Angelina V.; Sohn, Christine J.; Prunkard, Donna E.; Yakelis, Neal A.; Freeman, Jon O.

<P>Botryococcene is a branched triterpene produced by the algae <I>Botryococcus braunii</I>. Hydrocracking botryococcene yields a variety of combustible fuels such as gasoline and jet fuel. Engineering host systems and proteins involved in the biosynthesis of botryococcene to optimize production is of interest given these applications. The current study investigates the use of a diaryltetrazole based screen that undergoes a photoclick reaction with terminal alkenes, such as the branched terminal alkene present on botryococcene, to yield a fluorescent product. Host <I>E. coli</I> systems were established to produce botryococcene, squalene, and no triterpene to serve as a control. Cells were incubated with tetrazole and briefly irradiated with UV light to initiate the photoclick reaction. It was found that the botryococcene producing cells yielded observable fluorescence while the squalene and control cells had negligible fluorescence turn-on activity. Fluorescence-activated cell sorting (FACS) was subsequently used to identify and sort botryococcene producing <I>E. coli</I> from a mixture of control and squalene producing cells.</P><P>Graphic Abstract</P><P>Botryococcene is a branched triterpene produced by the algae <I>Botryococcus braunii</I>.<BR/><IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=c8ob02589d'/><BR/></P>

2019

The Royal Society of Chemistry

1477-0520

1477-0539

17

12

pp.3195-3201

10.1039/c8ob02589d

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART96201788