Synthetic Protein Scaffolds for Biosynthetic Pathway Colocalization on Lipid Droplet Membranes

ACS Synthetic Biology
Jyun-Liang LinIan Wheeldon

Abstract

Eukaryotic biochemistry is organized throughout the cell in and on membrane-bound organelles. When engineering metabolic pathways this organization is often lost, resulting in flux imbalance and a loss of kinetic advantages from enzyme colocalization and substrate channeling. Here, we develop a protein-based scaffold for colocalizing multienzyme pathways on the membranes of intracellular lipid droplets. Scaffolds based on the plant lipid droplet protein oleosin and cohesin-dockerin interaction pairs recruited upstream enzymes in yeast ester biosynthesis to the native localization of the terminal reaction step, alcohol-O-acetyltransferase (Atf1). The native localization is necessary for high activity and pathway assembly in close proximity to Atf1 increased pathway flux. Screening a library of scaffold variants further showed that pathway structure can alter catalysis and revealed an optimized scaffold and pathway expression levels that produced ethyl acetate at a rate nearly 2-fold greater than unstructured pathways. This strategy should prove useful in spatially organizing other metabolic pathways with key lipid droplet-localized and membrane-bound reaction steps.

References

Feb 1, 1989·Applied and Environmental Microbiology·E PostmaJ P Van Dijken
Feb 18, 1997·Proceedings of the National Academy of Sciences of the United States of America·G BarbaC Bréchot
Oct 21, 1999·Methods : a Companion to Methods in Enzymology·H O Spivey, J Ovádi
May 10, 2002·Nucleic Acids Research·David M Hoover, Jacek Lubkowski
Oct 25, 2003·Proceedings of the National Academy of Sciences of the United States of America·Albert SickmannChris Meisinger
Nov 19, 2003·Proceedings of the National Academy of Sciences of the United States of America·Ana L CarvalhoCarlos M G A Fontes
Feb 11, 2004·Biochemistry·Gerwald Jogl, Liang Tong
Sep 4, 2007·Trends in Biochemical Sciences·David W Piston, Gert-Jan Kremers
Feb 7, 2008·Annual Review of Plant Biology·Jörg Ziegler, Peter J Facchini
May 15, 2008·Journal of Cell Science·John K ZehmerRichard G W Anderson
Aug 22, 2008·Proceedings of the National Academy of Sciences of the United States of America·Jarrett J AdamsSteven P Smith
Aug 30, 2008·Current Opinion in Biotechnology·Robert J ConradoMatthew P DeLisa
Aug 4, 2009·Nature Biotechnology·John E DueberJay D Keasling
Nov 19, 2009·Proceedings of the National Academy of Sciences of the United States of America·Ella R Hinson, Peter Cresswell
Feb 2, 2010·Metabolic Engineering·Tae Seok MoonKristala L Jones Prather
May 11, 2010·The Journal of Physical Chemistry Letters·Patricia BaulerJ Andrew McCammon
Jun 28, 2011·Science·Camille J DelebecqueFaisal A Aldaye
Sep 29, 2011·Metabolic Engineering·Hanson LeeJohn E Dueber
Oct 25, 2011·Nucleic Acids Research·Robert J ConradoMatthew P DeLisa
May 19, 2012·Nature Chemical Biology·Jeong Wook LeeSang Yup Lee
Jul 24, 2012·Angewandte Chemie·Chun YouY-H Percival Zhang
Nov 20, 2012·Applied and Environmental Microbiology·Jun Hyoung LeeSun Chang Kim
Apr 10, 2013·Biotechnology and Bioengineering·Jang-Mi BaekMin-Kyu Oh
May 10, 2013·ACS Synthetic Biology·Shen-Long TsaiWilfred Chen
Aug 15, 2013·Journal of Cell Science·Samuel BouvetCatherine L Jackson
Sep 3, 2013·Methods in Molecular Biology·Zengyi Shao, Huimin Zhao
Sep 7, 2013·ACS Nano·Ofer Idan, Henry Hess
Mar 13, 2014·Nature Chemical Biology·Gabriel M RodriguezShota Atsumi
Sep 30, 2014·Nature Biotechnology·Michele CastellanaNed S Wingreen
Oct 5, 2014·Metabolic Engineering·Donovan S Layton, Cong T Trinh
Dec 3, 2014·Metabolic Engineering·Yi-Shu TaiKechun Zhang
Mar 24, 2016·Nature Chemistry·Ian WheeldonMatthew Sigman
Aug 4, 2016·ACS Synthetic Biology·Cameron MyhrvoldPamela A Silver

❮ Previous
Next ❯

Citations

Oct 17, 2018·Natural Product Reports·Camilla KnudsenTomas Laursen
Aug 31, 2017·Nature Chemical Biology·Sarah K Hammer, José L Avalos
Apr 1, 2020·Metabolites·Xueqin LvLong Liu
Apr 3, 2020·Chemical Communications : Chem Comm·Samuel LimDouglas S Clark
Oct 20, 2018·Frontiers in Bioengineering and Biotechnology·Leonardo Martins-SantanaRafael Silva-Rocha
Aug 2, 2018·Applied Microbiology and Biotechnology·Sarah Schmidt-DannertClaudia Schmidt-Dannert
May 16, 2019·Nature Chemical Biology·Evan M ZhaoJared E Toettcher
Feb 27, 2021·Frontiers in Bioengineering and Biotechnology·Hannah C YocumNancy A Da Silva
Oct 9, 2019·Current Opinion in Biotechnology·Stephen P LillingtonMichelle A O'Malley
Feb 26, 2021·Biotechnology Advances·Hao DongPing Wang
Nov 2, 2019·Current Opinion in Biotechnology·Toshihiro Obata
Dec 15, 2018·Current Opinion in Chemical Biology·Claudia Schmid-Dannert, Fernando López-Gallego
May 14, 2021·Journal of Agricultural and Food Chemistry·Jing WuXun Li
Jun 10, 2021·Physical Chemistry Chemical Physics : PCCP·Yan Xie, Scott Calabrese Barton
May 7, 2020·Bioconjugate Chemistry·Lenne J M LemmensLuc Brunsveld
Mar 20, 2019·ACS Synthetic Biology·James B Y H BehrendorffMathias Pribil

❮ Previous
Next ❯

Related Concepts

Related Feeds

Biomaterial Engineering

Advances in biomaterial engineering have permitted the development of sophisticated drug-releasing materials with a biomimetic 3D support that allow a better control of the microenvironment of transplanted cells. Here is the latest research.