Improving cell-free protein synthesis through genome engineering of Escherichia coli lacking release factor 1

Chembiochem : a European Journal of Chemical Biology
Seok Hoon HongMichael C Jewett

Abstract

Site-specific incorporation of non-standard amino acids (NSAAs) into proteins opens the way to novel biological insights and applications in biotechnology. Here, we describe the development of a high yielding cell-free protein synthesis (CFPS) platform for NSAA incorporation from crude extracts of genomically recoded Escherichia coli lacking release factor 1. We used genome engineering to construct synthetic organisms that, upon cell lysis, lead to improved extract performance. We targeted five potential negative effectors to be disabled: the nuclease genes rna, rnb, csdA, mazF, and endA. Using our most productive extract from strain MCJ.559 (csdA(-) endA(-)), we synthesized 550±40 μg mL(-1) of modified superfolder green fluorescent protein containing p-acetyl-L-phenylalanine. This yield was increased to ∼1300 μg mL(-1) when using a semicontinuous method. Our work has implications for using whole genome editing for CFPS strain development, expanding the chemistry of biological systems, and cell-free synthetic biology.

References

Sep 18, 1997·Proceedings of the National Academy of Sciences of the United States of America·D R LiuP G Schultz
Feb 19, 2002·Chemical Reviews·Ronald T. Raines
Aug 10, 2002·Journal of Bacteriology·Sidney R Kushner
Jan 1, 1965·Journal of Molecular Biology·G BERMARDO, C CORDONNIER
Mar 10, 2004·Biotechnology and Bioengineering·Michael C Jewett, James R Swartz
Jul 17, 2004·Metabolic Engineering·Nathalie Michel-ReydelletJames Swartz
Jul 23, 2004·Methods in Molecular Biology·James R SwartzKim A Woodrow
Jan 7, 2005·Biotechnology and Bioengineering·James Zawada, James Swartz
Apr 7, 2005·FEBS Letters·Dmitry E AgafonovMathias Sprinzl
Jun 7, 2005·Biotechnology and Bioengineering·Alexei M Voloshin, James R Swartz
Oct 29, 2005·Journal of Molecular Microbiology and Biotechnology·Nathalie Michel-ReydelletJames Swartz
Jan 31, 2006·Journal of Biotechnology·Kara A Calhoun, James R Swartz
Dec 26, 2006·Bioorganic & Medicinal Chemistry Letters·Shinsuke SandoYasuhiro Aoyama
Sep 11, 2008·Biotechnology and Bioengineering·Aaron R Goerke, James R Swartz
Apr 14, 2009·Nature Methods·Daniel G GibsonHamilton O Smith
Mar 24, 2010·Annual Review of Biochemistry·Chang C Liu, Peter G Schultz
Jun 24, 2010·Nucleic Acids Research·Randall A Hughes, Andrew D Ellington
Aug 13, 2010·Nucleic Acids Research·Takahito MukaiKensaku Sakamoto
Dec 18, 2010·Journal of the American Chemical Society·Isaac N UgwumbaColin J Jackson
Feb 2, 2011·Biochemistry·Douglas D YoungPeter G Schultz
Sep 20, 2011·Nature Chemical Biology·David B F JohnsonLei Wang
Sep 29, 2011·Metabolic Engineering·C Eric Hodgman, Michael C Jewett
Oct 20, 2011·Biotechnology Advances·Erik D CarlsonMichael C Jewett
Jan 13, 2012·Current Opinion in Biotechnology·Michael Georg Hoesl, Nediljko Budisa
Feb 1, 2012·Journal of the American Chemical Society·Yane-Shih WangWenshe R Liu
Feb 2, 2012·Biochemical and Biophysical Research Communications·Kiyoshi OzawaGottfried Otting
Feb 3, 2012·Angewandte Chemie·Karin V LoschaGottfried Otting
Jun 6, 2012·ACS Chemical Biology·David B F JohnsonLei Wang
Aug 7, 2012·Nature Chemical Biology·Ambra BiancoJason W Chin
Sep 7, 2012·Angewandte Chemie·Feng WangPeter G Schultz
Sep 19, 2012·Proceedings of the National Academy of Sciences of the United States of America·Jun Y AxupPeter G Schultz
Mar 28, 2013·Angewandte Chemie·Sangsik LeeHee-Sung Park
Apr 5, 2013·Angewandte Chemie·Abhishek ChatterjeePeter G Schultz
Jan 20, 2012·ACS Synthetic Biology·Jonghyeon Shin, Vincent Noireaux
May 15, 2013·ACS Chemical Biology·Wei NiuJiantao Guo
Oct 19, 2013·Science·Marc J LajoieFarren J Isaacs

❮ Previous
Next ❯

Citations

Jun 10, 2015·FEBS Letters·Mark J AndersonMichael C Jewett
Jun 18, 2015·Bioconjugate Chemistry·Dafni MoatsouRachel K O'Reilly
Oct 3, 2015·Metabolic Engineering·Jennifer E Kay, Michael C Jewett
Jul 18, 2015·Current Opinion in Chemical Biology·Benjamin J Des SoyeMichael C Jewett
Mar 22, 2016·Metabolic Engineering·Ashty S Karim, Michael C Jewett
Sep 10, 2015·Nature Communications·Javin P OzaMichael C Jewett
Jan 13, 2016·Biotechnology and Bioengineering·Rui Gan, Michael C Jewett
Jan 24, 2017·Biotechnology and Bioengineering·Jian LiMichael C Jewett
Feb 7, 2020·International Journal of Molecular Sciences·Sviatlana SmolskayaYaroslav A Andreev
Jul 13, 2016·Expert Review of Proteomics·Benjamin P MohrMitchel J Doktycz
Jun 6, 2019·Methods and Protocols·Nicole E GregorioJavin P Oza
Nov 5, 2019·Frontiers in Bioengineering and Biotechnology·Khushal KhambhatiVijai Singh
Nov 30, 2019·Nature Reviews. Genetics·Adam D SilvermanMichael C Jewett
Nov 30, 2019·Nucleic Acids Research·Michael J HammerlingMichael C Jewett
Aug 15, 2020·Frontiers in Bioengineering and Biotechnology·Yang WuHao Qi
Apr 9, 2020·Frontiers in Bioengineering and Biotechnology·Nadanai LaohakunakornSebastian J Maerkl
Feb 8, 2019·Critical Reviews in Biotechnology·Benjamin D TrumpIgor Linkov
Jun 21, 2020·Nature Communications·Luis E Contreras-LlanoCheemeng Tan
Oct 16, 2016·Cold Spring Harbor Perspectives in Biology·Jessica G PerezMichael C Jewett
Oct 23, 2020·Journal of Industrial Microbiology & Biotechnology·Jasmine HersheweMichael C Jewett
Sep 26, 2017·New Biotechnology·Daniel FoshagFrank Bernhard
Aug 11, 2020·Synthetic and Systems Biotechnology·Stephanie D ColeMatthew W Lux
Apr 6, 2016·Synthetic and Systems Biotechnology·Jennifer A SchoborgMichael C Jewett
Jun 8, 2021·Frontiers in Bioengineering and Biotechnology·Xinjie Chen, Yuan Lu
Aug 15, 2018·ACS Synthetic Biology·Benjamin J Des SoyeMichael C Jewett
Oct 26, 2018·ACS Synthetic Biology·Filippo CascheraMichael C Jewett
Jan 25, 2020·Journal of Agricultural and Food Chemistry·Fu-Xing NiuJian-Zhong Liu
Sep 10, 2021·Frontiers in Bioengineering and Biotechnology·Anibal ArceFernán Federici

❮ Previous
Next ❯

Related Concepts

Related Feeds

CZI Human Cell Atlas Seed Network

The aim of the Human Cell Atlas (HCA) is to build reference maps of all human cells in order to enhance our understanding of health and disease. The Seed Networks for the HCA project aims to bring together collaborators with different areas of expertise in order to facilitate the development of the HCA. Find the latest research from members of the HCA Seed Networks here.

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.