Development of Next Generation Synthetic Biology Tools for Use in Streptomyces venezuelae.
Streptomyces have a rich history as producers of important natural products and this genus of bacteria has recently garnered attention for its potential applications in the broader context of synthetic biology. However, the dearth of genetic tools available to control and monitor protein production precludes rapid and predictable metabolic engineering that is possible in hosts such as Escherichia coli or Saccharomyces cerevisiae. In an effort to improve genetic tools for Streptomyces venezuelae, we developed a suite of standardized, orthogonal integration vectors and an improved method to monitor protein production in this host. These tools were applied to characterize heterologous promoters and various attB chromosomal integration sites. A final study leveraged the characterized toolset to demonstrate its use in producing the biofuel precursor bisabolene using a chromosomally integrated expression system. These tools advance S. venezuelae to be a practical host for future metabolic engineering efforts.
Joint Bioenergy Institute , 5885 Hollis Street, Emeryville, California 94608, United States.
Ryan M
RM
Phelan
Joint Bioenergy Institute , 5885 Hollis Street, Emeryville, California 94608, United States.
Daniel
D
Sachs
Joint Bioenergy Institute , 5885 Hollis Street, Emeryville, California 94608, United States.
Shayne J
SJ
Petkiewicz
Joint Bioenergy Institute , 5885 Hollis Street, Emeryville, California 94608, United States.
Jesus F
JF
Barajas
Jacquelyn M
JM
Blake-Hedges
Mitchell G
MG
Thompson
Joint Bioenergy Institute , 5885 Hollis Street, Emeryville, California 94608, United States.
Amanda
A
Reider Apel
Department of Biology, Miami University , 212 Pearson Hall, Oxford, Ohio 45046, United States.
Blake J
BJ
Rasor
Leonard
L
Katz
Joint Bioenergy Institute , 5885 Hollis Street, Emeryville, California 94608, United States.|Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark , Kogle Allé, DK2970-Hørsholm, Denmark.
Jay D
JD
Keasling
D056804:Biofuels; D017930:Genes, Reporter; D005822:Genetic Vectors; D008164:Luminescent Proteins / Q000235:genetics; D060847:Metabolic Engineering / Q000379:methods; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D012717:Sesquiterpenes / Q000737:chemistry / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*; D058615:Synthetic Biology
D056804:Biofuels; D008164:Luminescent Proteins; D012717:Sesquiterpenes
bisabolene; fluorescent protein; promoter; streptomyces