Streptomyces venezuelae TX-TL - a next generation cell-free synthetic biology tool
File(s)Moore_et_al-2017-Biotechnology_Journal.pdf (1.21 MB)
Published version
Author(s)
Moore, SJ
Lai, HE
Needham, H
Polizzi, KM
Freemont, PS
Type
Journal Article
Abstract
Streptomyces venezuelae is a promising chassis in synthetic biology for fine chemical and secondary metabolite pathway engineering. The potential of S. venezuelae could be further realized by expanding its capability with the introduction of its own in vitro transcription-translation (TX-TL) system. TX-TL is a fast and expanding technology for bottom-up design of complex gene expression tools, biosensors and protein manufacturing. Herein, we introduce a S. venezuelae TX-TL platform by reporting a streamlined protocol for cell-extract preparation, demonstrating high-yield synthesis of a codon-optimized sfGFP reporter and the prototyping of a synthetic tetracycline-inducible promoter in S. venezuelae TX-TL based on the tetO-TetR repressor system. The aim of this system is to provide a host for the homologous production of exotic enzymes from Actinobacteria secondary metabolism in vitro. As an example, the authors demonstrate the soluble synthesis of a selection of enzymes (12-70 kDa) from the Streptomyces rimosus oxytetracycline pathway.
Date Issued
2017-02-24
Date Acceptance
2017-01-30
Citation
Biotechnology Journal, 2017, 12 (4)
ISSN
1860-7314
Publisher
Wiley-VCH Verlag
Journal / Book Title
Biotechnology Journal
Volume
12
Issue
4
Copyright Statement
© 2017 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. This is an open access article under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
License URL
Identifier
http://www.ncbi.nlm.nih.gov/pubmed/28139884
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Biotechnology & Applied Microbiology
Biochemistry & Molecular Biology
Cell-free
Gene expression
Streptomyces
Synthetic biology
Tetracycline
FREE PROTEIN-SYNTHESIS
BIOSYNTHESIS
TRANSLATION
PATHWAYS
RESOURCE
ENZYMES
Actinobacteria
Cell-Free System
Escherichia coli
Gene Expression Regulation, Bacterial
Metabolic Networks and Pathways
Oxytetracycline
Promoter Regions, Genetic
Protein Biosynthesis
Synthetic Biology
Transcription, Genetic
1002 Environmental Biotechnology
1003 Industrial Biotechnology
1004 Medical Biotechnology
Biotechnology
Publication Status
Published
Coverage Spatial
Germany
Article Number
1600678