A-94964, Novel Inhibitor of Bacterial Translocase I, Produced by Streptomyces sp. SANK 60404

Abstract

In our screening for translocase I inhibitors, we found the novel nucleoside antibiotic, A-94964 in the culture broth of Streptomyces sp. SANK 60404. The structure of A-94964 was elucidated primarily by various NMR studies, including a 1 H-31 P HMBC experiment. A-94964 has a unique structure which possesses a nucleoside moiety and an N-acylglucosamine moiety connected via a phosphate.

A comprehensive review of glycosylated bacterial natural products

“…888–890 A bacterial translocase inhibitor, A-94964A contains two unidentified sugars, 2′-deoxy-2′-aminohexosyl-1-phosphate ( 234 ) and a hexose ( 334 ), attached to a riboocturonic acid ( 233 ) appended to N -1 of uracil. 891,892 …”

Identification and Characterization of Bacterial Diterpene Cyclases that Synthesize the Cembrane Skeleton

“…SANK 60404 was a gift from Daiichi Sankyo (Tokyo, Japan), which had previously isolated the strain from a soil sample collected in Okinawa, Japan. [ 15 , 16 ] E. coli DH5α and E. coli BL21(DE3) (Takara Bio, Tokyo, Japan) were used for plasmid cloning and recombinant protein expression, respectively. The pT7Blue T-Vector (Novagen/Merck Millipore) and the pHis8 vector[27] were used to clone PCR products and express proteins, respectively.…”

“…SANK 60404, which was not previously known to be a terpenoid producer. [ 15 , 16 ] To mine these diterpene cyclases from the bacterium, we directed our attention to genes in close proximity to a gene encoding a GGDP synthase, which is indispensable for diterpene production. There are two reasons for this choice of approach: 1) the bacterial diterpene cyclases mentioned above exhibit only a low level of overall sequence similarity, whereas the GGDP synthases from these bacteria display more than 30 % identity, [ 8 , 10 , 13 ] and 2) a GGDP synthase gene is located near a diterpene cyclase gene in the genomes of these bacteria (Figure S2).…”

A novel assay of bacterial peptidoglycan synthesis for natural product screening

“…We applied this method to high-throughput screening of natural products and discovered novel translocase I inhibitors. [13][14][15][16][17][32][33][34][35] As the assay is sensitive and robust enough, we tried to measure MurF activity by coupling with this reaction (Figure 1). For that purpose, we first established a large-scale fermentation protocol for UDPMurNAc-tripeptide to prepare the labeled substrate UDP-MurNAcdansyltripeptide.…”