PMID- 34894206
OWN - NLM
STAT- MEDLINE
DCOM- 20220407
LR  - 20231130
IS  - 1860-7314 (Electronic)
IS  - 1860-6768 (Linking)
VI  - 17
IP  - 4
DP  - 2022 Apr
TI  - Improving genomically recoded Escherichia coli to produce proteins containing 
      non-canonical amino acids.
PG  - e2100330
LID - 10.1002/biot.202100330 [doi]
AB  - A genomically recoded Escherichia coli strain that lacks all amber codons and 
      release factor 1 (C321.∆A) enables efficient genetic encoding of chemically 
      diverse non-canonical amino acids (ncAAs) into proteins. While C321.∆A has opened 
      new opportunities in chemical and synthetic biology, this strain has not been 
      optimized for protein production, limiting its utility in widespread industrial 
      and academic applications. To address this limitation, the construction of a 
      series of genomically recoded organisms that are optimized for cellular protein 
      production is described. It is demonstrated that the functional deactivation of 
      nucleases (e.g., rne, endA) and proteases (e.g., lon) increases production of 
      wild-type superfolder green fluorescent protein (sfGFP) and sfGFP containing two 
      ncAAs up to  approximately 5-fold. Additionally, a genomic IPTG-inducible T7 RNA polymerase 
      (T7RNAP) cassette into these strains is introduced. Using an optimized platform, 
      the ability to introduce two identical N(6) -(propargyloxycarbonyl)-(L) -Lysine 
      residues site specifically into sfGFP with a 17-fold improvement in production 
      relative to the parent strain is demonstrated. The authors envision that their 
      library of organisms will provide the community with multiple options for 
      increased expression of proteins with new and diverse chemistries.
CI  - (c) 2021 Wiley-VCH GmbH.
FAU - Perez, Jessica G
AU  - Perez JG
AD  - Department of Chemical and Biological Engineering, Northwestern University, 
      Evanston, Illinois, USA.
AD  - Chemistry of Life Processes Institute, Northwestern University, Evanston, 
      Illinois, USA.
FAU - Carlson, Erik D
AU  - Carlson ED
AD  - Department of Chemical and Biological Engineering, Northwestern University, 
      Evanston, Illinois, USA.
AD  - Chemistry of Life Processes Institute, Northwestern University, Evanston, 
      Illinois, USA.
FAU - Weisser, Oliver
AU  - Weisser O
AD  - Department of Chemical and Biological Engineering, Northwestern University, 
      Evanston, Illinois, USA.
AD  - Chemistry of Life Processes Institute, Northwestern University, Evanston, 
      Illinois, USA.
FAU - Kofman, Camila
AU  - Kofman C
AD  - Department of Chemical and Biological Engineering, Northwestern University, 
      Evanston, Illinois, USA.
AD  - Chemistry of Life Processes Institute, Northwestern University, Evanston, 
      Illinois, USA.
FAU - Seki, Kosuke
AU  - Seki K
AD  - Department of Chemical and Biological Engineering, Northwestern University, 
      Evanston, Illinois, USA.
AD  - Chemistry of Life Processes Institute, Northwestern University, Evanston, 
      Illinois, USA.
FAU - Des Soye, Benjamin J
AU  - Des Soye BJ
AD  - Department of Chemical and Biological Engineering, Northwestern University, 
      Evanston, Illinois, USA.
AD  - Chemistry of Life Processes Institute, Northwestern University, Evanston, 
      Illinois, USA.
FAU - Karim, Ashty S
AU  - Karim AS
AUID- ORCID: 0000-0002-5789-7715
AD  - Department of Chemical and Biological Engineering, Northwestern University, 
      Evanston, Illinois, USA.
AD  - Chemistry of Life Processes Institute, Northwestern University, Evanston, 
      Illinois, USA.
FAU - Jewett, Michael C
AU  - Jewett MC
AUID- ORCID: 0000-0003-2948-6211
AD  - Department of Chemical and Biological Engineering, Northwestern University, 
      Evanston, Illinois, USA.
AD  - Chemistry of Life Processes Institute, Northwestern University, Evanston, 
      Illinois, USA.
AD  - Center for Synthetic Biology, Northwestern University, Evanston, Illinois, USA.
AD  - Simpson Querrey Institute, Northwestern University, Chicago, Illinois, USA.
LA  - eng
GR  - T32 GM105538/GM/NIGMS NIH HHS/United States
PT  - Journal Article
DEP - 20211229
PL  - Germany
TA  - Biotechnol J
JT  - Biotechnology journal
JID - 101265833
RN  - 0 (Amino Acids)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - *Amino Acids/metabolism
MH  - *Escherichia coli/genetics/metabolism
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - Synthetic Biology
OTO - NOTNLM
OT  - amber suppression
OT  - genomically recoded organism
OT  - non-canonical amino acids
OT  - orthogonal translational system
OT  - protein production
EDAT- 2021/12/12 06:00
MHDA- 2022/04/08 06:00
CRDT- 2021/12/11 08:38
PHST- 2021/12/08 00:00 [revised]
PHST- 2021/06/26 00:00 [received]
PHST- 2021/12/09 00:00 [accepted]
PHST- 2021/12/12 06:00 [pubmed]
PHST- 2022/04/08 06:00 [medline]
PHST- 2021/12/11 08:38 [entrez]
AID - 10.1002/biot.202100330 [doi]
PST - ppublish
SO  - Biotechnol J. 2022 Apr;17(4):e2100330. doi: 10.1002/biot.202100330. Epub 2021 Dec 
      29.