Affibody scaffolds improve sesquiterpene production in Saccharomyces cerevisiae

Publication: Research - peer-reviewLetter – Annual report year: 2016

DOI

View graph of relations

Enzyme fusions have been widely used as a tool in metabolic engineering to increase pathway efficiency by reducing substrate loss and accumulation of toxic intermediates. Alternatively, enzymes can be co-localized through attachment to a synthetic scaffold via non-covalent interactions. Here we describe the use of affibodies for enzyme tagging and scaffolding. The scaffolding is based on the recognition of affibodies to their anti-idiotypic partners in vivo, and was first employed for co-localization of farnesyl diphosphate synthase and farnesene synthase in S. cerevisiae. Different parameters were modulated to improve the system, and the enzyme:scaffold ratio was most critical for its functionality. Ultimately, the yield of farnesene on glucose YSFar could be improved by 135 % in fed-batch cultivations using a 2-site affibody scaffold. The scaffolding strategy was then extended to a three-enzyme polyhydroxybutyrate (PHB) pathway, heterologously expressed in E. coli. Within a narrow range of enzyme and scaffold induction, the affibody tagging and scaffolding increased PHB production 7-fold. This work demonstrates how the versatile affibody can be used for metabolic engineering purposes.
Original languageEnglish
JournalA C S Synthetic Biology
Volume6
Issue number1
Pages (from-to)19-28
Number of pages10
ISSN2161-5063
DOIs
StatePublished - 2017
CitationsWeb of Science® Times Cited: 1

    Keywords

  • Affibodies, Biofuels, Isoprenoids, Metabolic engineering, PHB, Yeast
Download as:
Download as PDF
Select render style:
APAAuthorCBE/CSEHarvardMLAStandardVancouverShortLong
PDF
Download as HTML
Select render style:
APAAuthorCBE/CSEHarvardMLAStandardVancouverShortLong
HTML
Download as Word
Select render style:
APAAuthorCBE/CSEHarvardMLAStandardVancouverShortLong
Word

ID: 127386259