Komagataeibacter tool kit (KTK): a modular cloning system for multigene constructs and programmed protein secretion from cellulose producing bacteria
File(s)Manuscript File_Goosens_2021_ACS SB FINAL.docx (1.83 MB)
Accepted version
Author(s)
Type
Journal Article
Abstract
Bacteria proficient at producing cellulose are an attractive synthetic biology host for the emerging field of Engineered Living Materials (ELMs). Species from the Komagataeibacter genus produce high yields of pure cellulose materials in a short time with minimal resources, and pioneering work has shown that genetic engineering in these strains is possible and can be used to modify the material and its production. To accelerate synthetic biology progress in these bacteria, we introduce here the Komagataeibacter tool kit (KTK), a standardised modular cloning system based on Golden Gate DNA assembly that allows DNA parts to be combined to build complex multigene constructs expressed in bacteria from plasmids. Working in Komagataeibacter rhaeticus, we describe basic parts for this system, including promoters, fusion tags and reporter proteins, before showcasing how the assembly system enables more complex designs. Specifically, we use KTK cloning to reformat the Escherichia coli curli amyloid fibre system for functional expression in K. rhaeticus, and go on to modify it as a system for programming protein secretion from the cellulose producing bacteria. With this toolkit, we aim to accelerate modular synthetic biology in these bacteria, and enable more rapid progress in the emerging ELMs community.
Date Issued
2021-12-17
Date Acceptance
2021-11-01
Citation
ACS Synthetic Biology, 2021, 10 (12), pp.3422-3434
ISSN
2161-5063
Publisher
American Chemical Society
Start Page
3422
End Page
3434
Journal / Book Title
ACS Synthetic Biology
Volume
10
Issue
12
Copyright Statement
© 2021 American Chemical Society. This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Synth. Biol., after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acssynbio.1c00358
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Engineering and Physical Sciences Research Council
Engineering & Physical Science Research Council (EPSRC)
Commission of the European Communities
US Army (US)
Engineering & Physical Science Research Council (EPSRC)
Identifier
https://pubs.acs.org/doi/10.1021/acssynbio.1c00358
Grant Number
EP/M002306/1
EP/N026489/1
EP/N026489/1
820699
W911NF1810387
EP/S032215/1
Subjects
0304 Medicinal and Biomolecular Chemistry
0601 Biochemistry and Cell Biology
0903 Biomedical Engineering
Publication Status
Published
Date Publish Online
2021-11-12