Genome mining reveals a minimum gene set for the biosynthesis of 32-membered macrocyclic thiopeptides lactazoles.
Although >100 thiopeptides have been discovered, the number of validated gene clusters involved in their biosynthesis is lagging. We use genome mining to identify a silent thiopeptide biosynthetic gene cluster responsible for biosynthesis of lactazoles. Lactazoles are structurally unique thiopeptides with a 32-membered macrocycle and a 2-oxazolyl-6-thiazolyl pyridine core. We demonstrate that lactazoles originate from the simplest cluster, containing only six unidirectional genes (lazA to lazF). We show that lazC is involved in the macrocyclization process, leading to central pyridine moiety formation. Substitution of the endogenous promoter with a strong promoter results in an approximately 30-fold increase in lactazole A production and mutagenesis of lazC precursor gene in production of two analogs. Lactazoles do not exhibit antimicrobial activity but may modulate signaling cascades triggered by bone morphogenetic protein. Our approach facilitates the production of a more diverse set of thiopeptide structures, increasing the semisynthetic repertoire for use in drug development.
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
Shohei
S
Hayashi
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
Taro
T
Ozaki
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
Shumpei
S
Asamizu
Kitasato Institute for Life Sciences, Kitasato University, Sagamihara 228-8555, Japan.
Haruo
H
Ikeda
The Kitasato Institute, Kitasato University, Tokyo 108-8642, Japan.
Satoshi
S
Ōmura
Biotechnology Research Center, Toyama Prefectural University, Imizu 939-0398, Japan.
Naoya
N
Oku
Biotechnology Research Center, Toyama Prefectural University, Imizu 939-0398, Japan.
Yasuhiro
Y
Igarashi
Graduate School of Pharmaceutical Sciences, Kitasato University, Tokyo 108-8641, Japan.
Hiroshi
H
Tomoda
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan; Biotechnology Research Center, Toyama Prefectural University, Imizu 939-0398, Japan. Electronic address: aonaka@mail.ecc.u-tokyo.ac.jp.
Hiroyasu
H
Onaka
D053898:Biosynthetic Pathways / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D005818:Genetic Engineering; D016680:Genome, Bacterial / Q000235:genetics*; D047028:Macrocyclic Compounds / Q000737:chemistry / Q000378:metabolism*; D015394:Molecular Structure; D005810:Multigene Family / Q000235:genetics; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D010456:Peptides, Cyclic / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D047028:Macrocyclic Compounds; D010455:Peptides; D010456:Peptides, Cyclic; D013438:Sulfhydryl Compounds; D013844:Thiazoles