5-Hydroxymethylblasticidin S and blasticidin S from Streptomyces setonii culture A83094.

Abstract

In our preceding paperx) we described the isolation and structure elucidation of antibiotic A83094A(16-deethylindanomycin) from the biomass of Streptomyces setonii. The culture filtrate from a fermentation of this same organism contains two broad-spectrum antibiotics which were isolated and determined to be 5-hydroxymethylblasticidin S (A83094B) and blasticidin S (A83094C) as shown in Fig. 1.The flow diagram for the isolation of 5-hydroxymethylblasticidin S and blasticidin S as a complex is presented in Fig. 2. The antibiotic levels at each step were determined both by disc plate assay vs. Salmonella gallinarum and by HPLC.HPLCassays were run on a ywBondapak C18 column (3.9 x 300 mm) using a mobile phase ofCH3CN -H2O (4 : 96) containing 1 % NH4OAc (w/v) and UV detection at 225 nm.Separation of the complex into individual antibiotics was accomplished using semi-preparative HPLC. The reversed phase chromatography system consisted of a M6000 pump, /*Bondapak C18 column (9.8 x 300 mm), Model 490 variable wavelength detector.U6K man- and mobile phase (as described above for analytical HPLC) pumped at 4ml/minute. Eight separate chromatographic runs were performed as follows: 20 jul aqueous sample, containing 2 mgantibiotic mixture, was applied.16 ml of eluate was discarded; then fractions were collected at 20-second intervals and assayed by HPLC. Fractions containing >95% of each component were combined and lyophilized.Each lyophile was dissolved in 1.0 ml water and desalted by injection on a^Bondapak C18 column (3.9x300mm).After washing the column with 20ml water, the antibiotic was elutedwith CH3CN-H2O (50 : 50). The CH3CN was removed under reduced pressure and the remaining solution lyophilized to provide samples for physico-chemical analyses. The *H NMR of A83094B (500 MHz, DMSOd6) was compared to authentic blasticidin S and indicates that they are structurally similar (see Table 1). The differences are in the loss of 5-H in A83094B and the addition of a hydroxymethylene at <5H 4.21. A nuclear Overhauser effect (NOE) is observed between the new methylene and 6-H. The presence of a hydroxymethylene is consistent with the mass spectral data. Based on recently published data, the structure of A83094B is similar to Sch 366054).

Cytosylglucuronic acid synthase (cytosine: UDP-glucuronosyltransferase) from Streptomyces griseochromogenes, the first prokaryotic UDP-glucuronosyltransferase

“…Although BS is most potent as an antifungal agent, it also exhibits antibacterial (32), antiviral (16,17), and antitumor activities (39). Several other peptidyl nucleoside antibiotics have close structural similarities to BS (1,7,15,19,20,36,37); they all consist of a cytosine hexuronic nucleoside unit and a basic amino acid side chain.…”

“…As shown in Figure 2, SDS-PAGE of the purified CGA synthase from the last step (lanes 8 CGA synthase, isolated from S. griseochromogenes, catalyzes the formation of CGA from UDP-glucuronic acid and cytosine in the biosynthesis of the peptide-nucleoside antibiotic BS (14) and is also quite likely involved in the biosynthesis of a number of other known nucleoside antibiotics (1,7,15,19,20,36,37). Functionally, CGA synthase belongs to the family of UDPGTs; although UDP-GTs are common in mammalian metabolism and at least two have been reported for fungi (10,40), they have not been previously reported for a prokaryotic organism.…”

Detection, characterization, and phytotoxic activity of the nucleoside antibiotics, Blasticidin S and 5-Hydroxylmethyl-Blasticidin S

“…The physicochemical data for BI-S have already been reported in several papers (Otake et al 1966, Takeuchi et al 1958, Yonehara 1984; therefore, we report only those that supplement the reported data (Larsen et al 1989) for H-M-BI-S.…”

“…It acts against fungi and bacteria by inhibiting protein synthesis with a mechanism similar to Puromycin (Yonehara 1984). H-M-BI-S has recently been isolated together with B1-S from the culture broth of Streptomyces setonii (Larsen et al 1989).…”

Isolation of Bacillus atrophaeus MPL-01 from A Wild Boar and Characterization of Its Antifungal Activity

“…1960년대 이후 환경오염을 줄이고 식물병원성 진균에 의한 피해를 줄일 수 있는 화학 농약 대체 생물 학적 제제를 개발하려는 시도가 이루어져 blasticidin, kasugamycin 등이 실제 방제에 적용되었다. 몇몇의 이차대사물질은 독성 등의 이유로 실용화하지 못하고 있는 실정이다 (Larsen et al, 1989;Hotta, 1996). 이러한 문제점 때문에 방선균 이외의 균을 이용한 식물병 방제제 개발이 시도되었다.…”