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).

Structure of the New Spiroketal-Macrolide A82548A. --(KIRST, H. A.; LARSEN, S. H.; PASCHAL, J. W.; OCCOLOWITZ, J. L.; CREEMER, L. C.; RIOS STEINER, J. L.; LOBKOVSKY, E.; CLARDY, J.; J. Antibiot. 48 (1995) 9, 990-996; Lilly Res. Lab., Greenfield, IN 46140, USA; EN)

C179touchscreens, barcode scanners, digital cameras, and other forms of automation.Using these data, the Xtaldb system organizes experiments and overall target status information into projects and provides tools for data mining and statistical analysis of the crystallization data both on the project and database-wide levels. To test these tools, we used the system in the salvage of a group of ten targets that previously failed to produce a structure in the MCSG pipeline. To date, two structures have been solved and deposited in the PDB, and three others diffract natively: two to 2.7Å, and one to 3.6Å. Protein adsorption on solid surfaces has a wide range of applications [1]. The use of Grazing Incidence X-Ray techniques to investigate protein structure adsorbed on interfaces is a promising tool that may lead to the understanding of its function. In diazotroph microorganisms, GlnB of H. seropedicae signalizes levels of nitrogen for a series of proteins involved in the regulation of expression and activity of nitrogenase complex. The GlnB-HS structure was already determined by x-ray diffraction revealing a trimmer of (36kDa) [2].The subject of this investigation is to understand the interaction of protein GlnB-Hs, a globular protein, on Si (111) and Si(100) surfaces under different conditions of deposition. The spin coating technique [3] was used to obtain a uniform thin film. This experiment was conducted on a Huber six-circles diffractometer, at XRD2 beamline(LNLS-Brazil), with energy tuned to approximately 7Kev. The results were used to obtain information on protein layer assembly. The initial scattering profiles of standard -2 obtained in grazing incidence geometries showed signal of protein layers ordering corresponding to a d-spacing of 30 in Si(111) and 40 in Si(100) out of plane direction compatible with crystallographic data.[1] Gray J., Curr. Opi. in Struct. Biology, 2004, 14, 110. [2] Benelli E., Buck M., Polikarpov I., DeSouza E., Cruz L., Pedrosa F., Eur. J. Biochem., 2002, 269, 3296. [3] In the post human genome era, the focus has shifted from sequencing genomes to investigate the proteins that are encoded by the genomes. The structural genomics programs have different missions but they all share the fact, that they have put together a high throughput pipeline that make it cheaper, easier and faster to get from gene to the three dimensional structure of the encoded protein. In this pipeline there are several bottlenecks, but it is agreed in general that the most significant bottleneck is to get from the protein solution to protein crystals of diffraction quality.We have implemented the method of Multilayer Soft Lithography to produce Formulator chips [1] to address the problem of protein crystallization. Using the Formulator chip, the solubility behaviour of a protein can be experimental characterised using minute volumes of sample. The protein is screened against 3000 chemical conditions using less than 10 µL of purified protein sample. Subsequently 30 to 50 chemical conditions from this sparse...