Tokyo University of Agriculture and Technology

Institute of Global Innovation Research

グローバルイノベーション研究院

公開セミナー

Institute of Global Innovation Research Open Seminar

Chassis-Independent Recombinase Assisted GenomeEngineering Enables Rapid Activation of Secondary Metabolite
Biosynthetic Clusters in Undomesticated Bacteria
Dr. Yasuo Yoshikuni

Head DNA Synthesis Science Program,
The U.S. Department of Energy Joint Genome Institute,
Lawrence Berkeley National Laboratory, U.S.A.

Monday,Abstract

December

16 2019, 17:00 ~ 18:00

東京農工大学 小金井キャンパス 11号館 講義室 L1153
Lecture Room L1153, Building 11, Koganei Campus TUAT

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 productsʼ
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 highaccuracy 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 nativestrain 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 thatsystematic approaches based on CRAGE will be useful for discovering and identifying previously uncharacterized
metabolites.
References
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 ofmicrobial 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.

■共催 / Co-Organized by

グローバルイノベーション研究院 エネルギー分野 新垣チーム
Institute of Global Innovation “Energy” Arakaki Team
卓越大学院プログラム
Excellent Leader Development for Super Smart Society
by New Industry Creation and Diversity■お問合せ先／Contact
グローバルイノベーション研究院 工学研究院 新垣篤史
Institute of Global Innovation Research, Institute of Engineering,
Assoc. Prof. Atsushi Arakaki
Email: arakakia (ここに@を入れてください) cc.tuat.ac.jp
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