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Enhanced Production of Dopa-Incorporated Mussel Adhesive Protein Using Engineered Translational Machineries
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  • Ye Seul Jeong,
  • Byeongseon Yang,
  • Byungseop Yang,
  • Mincheol Shin,
  • Jihyoun Seong,
  • Hyung Joon Cha,
  • Inchan Kwon
Ye Seul Jeong
Gwangju Institute of Science and Technology
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Byeongseon Yang
POSTECH
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Byungseop Yang
Gwangju Institute of Science and Technology
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Mincheol Shin
POSTECH
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Jihyoun Seong
Gwangju Institute of Science and Technology
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Hyung Joon Cha
POSTECH
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Inchan Kwon
Gwangju Institute of Science and Technology
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Abstract

Mussel adhesive proteins (MAPs) have great potential as bioglues, in particular in wet conditions. Although in vivo residue-specific incorporation of 3,4-dihydroxyphenylalanine (Dopa) in tyrosine-auxotrophic Escherichia coli cells allows production of bioengineered MAPs (bMAPs), the low production yield hinders the practical application of bMAPs. Such low production yield of Dopa-incorporated bMAPs (Dopa-bMAPs) was known to be caused by low translational activity of a noncanonical amino acid, Dopa, in E. coli cells. Herein, in order to enhance the production yield of Dopa-bMAPs, we investigated the coexpression of Dopa-recognizing tyrosyl-tRNA synthetases (TyrRSs). In order to use the Dopa-specific Methanococcus jannascii TyrRS (MjTyrRS-Dopa), we altered the anti-codon of tyrosyl-tRNA amber suppressor into AUA (MjtRNATyrAUA) to recgonize a tyrosine codon (MjtRNATyrAUA). Co-overexpression of MjTyrRS-Dopa and MjtRNATyrAUA increased the production yield of Dopa-MAP by 57%. Similarly, overexpression of E. coli TyrRS (EcTyrRS) led to a 72% higher production yield of Dopa-incorporated bMAP. Even with coexpression of Dopa-recognizing TyrRSs, Dopa-bMAPs have a high Dopa incorporation yield (over 90%) compared to Dopa-bMAPs prepared without any coexpression of TyrRS.

Peer review status:ACCEPTED

26 Dec 2019Submitted to Biotechnology and Bioengineering
27 Dec 2019Submission Checks Completed
27 Dec 2019Assigned to Editor
07 Jan 2020Reviewer(s) Assigned
04 Feb 2020Review(s) Completed, Editorial Evaluation Pending
04 Feb 2020Editorial Decision: Revise Minor
09 Feb 20201st Revision Received
10 Feb 2020Assigned to Editor
10 Feb 2020Submission Checks Completed
11 Feb 2020Reviewer(s) Assigned
29 Feb 2020Review(s) Completed, Editorial Evaluation Pending
29 Feb 2020Editorial Decision: Revise Minor
07 Mar 20202nd Revision Received
07 Mar 2020Submission Checks Completed
07 Mar 2020Assigned to Editor
12 Mar 2020Reviewer(s) Assigned
13 Mar 2020Review(s) Completed, Editorial Evaluation Pending
13 Mar 2020Editorial Decision: Revise Minor
16 Mar 20203rd Revision Received
16 Mar 2020Submission Checks Completed
16 Mar 2020Assigned to Editor
18 Mar 2020Review(s) Completed, Editorial Evaluation Pending
18 Mar 2020Editorial Decision: Accept