Acetone) was added to the cultures. The progress of conversion wasAcetone) was added towards the

Acetone) was added to the cultures. The progress of conversion was
Acetone) was added towards the cultures. The progress of conversion was monitored by TLC. Just after biotransformations, the metabolites and remaining substrate had been extracted with TXA2/TP Inhibitor Purity & Documentation methylene chloride. The organic solutions had been dried with anhydrous magnesium sulphate, filtered, concentrated in vacuo and analysed by GC. Within the analytical scale biotransformations making use of selected strains, 0.2 g of 1 dissolved in two ml of acetone was equally distributed amongst flasks with fungal cultures. The reactions have been carried out below the exact same situations as in screening tests and continued till the substrate was metabolized. The progress of conversion was monitored by TLC. When the transformation completed, mycelia and broth were extracted three occasions with methylene chloride. The organic extracts have been combined, dried more than anhydrous magnesium sulphate and filtered, and the solvent was evaporated in vacuo. These crude extracts had been analysed by TLC and GC and then chromatographed on a column of silica gel. Merchandise analysis TLC of crude extracts was carried out with Merck Kieselgel 60 F254 plates, visualized by spraying them using a mixture of methanol in concentrated sulphuric acid (1:1 v: v) and heating to 120 till the colours developed. Metabolites obtained in the analytical transformations were separated by column chromatography on silica gel 60 (23000 mesh) eluting with all the very same eluent as for TLC. GC evaluation was performed employing Hewlett Packard 5890A Series II GC instrument (FID, carrier gas H2 at flow price of 2 ml min-1) with DB-5MS column (crosslinked phenyl methyl siloxane, 30 m 9 0.32 mm 9 0.25 lm). The applied temperature program was 220 1 min-1, gradient 4 min-1 to 280 and then 30 to 300 three min-1; injector and detector temperature were 300 (for L. sulphureus temperature plan was 215 1 min-1, gradient four min-1 to 280 after which 30 to 300 three min-1). MS analyses have been performed on Varian CP-3800/Saturn 2000 apparatus having a Zebron ZB-5 MSI (30 m 9 0.25 mm 9 0.25 lm) column. The following temperature program was made use of: 220 1 min-1, gradient five min-1 to 300 five min-1. The NMR spectra have been recorded on a Bruker AvanceTM 600 MHz2021 The Authors. Microbial Biotechnology published by Society for Applied Microbiology and John Wiley Sons Ltd., Microbial Biotechnology, 14, 2187Microbial transformations of 7-oxo-DHEA (62 mg; 31 mol.), identified 3b,17b-dihydroxy-androst-5en-7-one (two) (30 mg; 15 mol.), in addition to a new product characterized as 3b,16b-dihydroxy-androst-5-en-7,17dione (six) (57 mg; 27 mol., Rt = 19.four min). 3b,16b-Dihydroxy-androst-5-en-7,17-dione (6): white amorphous solid; 1H NMR (CD3OD, 600 MHz) d: 0.96 (3H, s, H-18), 1.27 (3H, s, H-19), three.14-3.18 (1H, m, H15a), 3.54.60 (1H, m, H-3a), 3.94 (1H, t, J = eight.5 Hz, H-16a), five.72 (1H, d, J = 1.7 Hz, H-6). 13C NMR (CD3OD, 151 MHz) d: 14.9 (CH3, C-18), 17.7 (CH3, C19), 21.4 (CH2, C-11), 31.9 (CH2, C-2), 32.1 (CH2, C12), 34.5 (CH2, C-15), 37.four (CH2, C-1), 39.9 (C, C-10), 41.1 (CH, C-14), 42.eight (CH2, C-4), 44.7 (CH, C-8), 48.two (C, C-13), 51.six (CH, C-9), 71.1 (CH, C-3), 75.four(CH, C16), 126.1 (CH, C-6), 169.six (C, C-5), 203.3 (C, C-7), 220.7 (C, C-17). EI-MS m/z 318.five [M]+(27), 290.4 (100), 192.5 (48), 91.5 (66), 77.four (33). Biotransformation with Fusicoccum amygdali AM258 7-Oxo-DHEA (0.two g) dissolved in 2 ml of acetone was evenly distributed among two flasks with 4 days old fungal cultures and MMP-1 Inhibitor list incubated for additional 7 days. The normal procedure gave extracts, which had been purified on silica gel. Elution with acetone:et.