Institute of Global Innovation Research
|Date||2019.12.16 (17 : 00 - 18：00)|
|Venue||Lecture Room L1153, Building 11, Koganei Campus, TUAT|
|Speaker / Topic||Dr. Yasuo Yoshikuni (Head DNA Synthesis Science Program, The U.S. Department of Energy Joint Genome Institute, Lawrence Berkeley National Laboratory, U.S.A.)
"Chassis-Independent Recombinase Assisted Genome Engineering Enables Rapid Activation of Secondary Metabolite Biosynthetic Clusters in Undomesticated Bacteria"
It is generally believed that exchange of secondary metabolite biosynthetic gene clusters (BGCs)
among closely related bacteria is an important driver of BGC evolution and diversification.
Applying this idea may help researchers efficiently connect many BGCs to their products and
characterize the products’ roles in various environments. However, existing genetic tools support
only a small fraction of these efforts. Here, we present the development of chassis-independent
recombinase-assisted genome engineering (CRAGE), which enables single-step integration of
large, complex BGC constructs directly into the chromosomes of diverse bacteria with high
accuracy and efficiency. To demonstrate the efficacy of CRAGE, we expressed three known and
six previously identified but experimentally elusive non-ribosomal peptide synthetase (NRPS)
and NRPS-polyketide synthase (PKS) hybrid BGCs from Photorhabdus luminescens in 25
diverse γ-Proteobacteria species. Successful activation of six BGCs identified 22 products for
which diversity and yield were greater when the BGCs were expressed in strains closely related
to the native strain than when they were expressed in either native or more distantly related
strains. Activation of these BGCs demonstrates the feasibility of exploiting their underlying
catalytic activity and plasticity, and provides evidence that systematic approaches based on
CRAGE will be useful for discovering and identifying previously uncharacterized metabolites.
1. Coates, R.C. et al. An integrated workflow for phenazine-modifying enzyme
characterization. J Ind Microbiol Biotechnol 45, 567-577 (2018).
2. Cole, B.J. et al. Genome-wide identification of bacterial plant colonization genes. PLoS
Biol 15, e2002860 (2017).
3. Ke, J. & Yoshikuni, Y. Multi-chassis engineering for heterologous production of
microbial natural products. Curr Opin Biotechnol 62, 88-97 (2019).
4. Shulse, C.N. et al. Engineered Root Bacteria Release Plant-Available Phosphate from
Phytate. Appl Environ Microbiol 85 (2019).
5. Wang, G. et al. CRAGE enables rapid activation of biosynthetic gene clusters in
undomesticated bacteria. Nat Microbiol (2019).
|Intended for||Everyone is welcome to attend.|
|Co-Organized by||Institute of Global Innovation “Energy” Arakaki Team
Excellent Leader Development for Super Smart Society by New Industry Creation and Diversity
|Contact||Institute of Global Innovation Research, Institute of Engineering
Assoc. Prof. Atsushi Arakaki
e-mail: arakakia (at) cc.tuat.ac.jp
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