In:
Frontiers in Bioengineering and Biotechnology, Frontiers Media SA, Vol. 10 ( 2022-9-16)
Abstract:
Cell-free gene expression (CFE) systems are an attractive tool for engineering within synthetic biology and for industrial production of high-value recombinant proteins. CFE reactions require a cell extract, energy system, amino acids, and DNA, to catalyse mRNA transcription and protein synthesis. To provide an amino acid source, CFE systems typically use a commercial standard, which is often proprietary. Herein we show that a range of common microbiology rich media (i.e., tryptone, peptone, yeast extract and casamino acids) unexpectedly provide an effective and low-cost amino acid source. We show that this approach is generalisable, by comparing batch variability and protein production in the following range of CFE systems: Escherichia coli (Rosetta ™ 2 (DE3), BL21(DE3)), Streptomyces venezuelae and Pichia pastoris . In all CFE systems, we show equivalent or increased protein synthesis capacity upon replacement of the commercial amino acid source. In conclusion, we suggest rich microbiology media provides a new amino acid source for CFE systems with potential broad use in synthetic biology and industrial biotechnology applications.
Type of Medium:
Online Resource
ISSN:
2296-4185
DOI:
10.3389/fbioe.2022.992708
DOI:
10.3389/fbioe.2022.992708.s001
Language:
Unknown
Publisher:
Frontiers Media SA
Publication Date:
2022
detail.hit.zdb_id:
2719493-0
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