Isolation of 2-amino-2-deoxy-D-mannitol from culture broth of Streptomyces species.

Agricultural and Biological Chemistry
1989.0

Abstract

Numerous papers have been published on bacterial polysaccharides containing amino sugars or aminouronic acids, and on oligosaccharides containing amino sugars in glycoproteins, glycosaminoglycans and glycolipids. The title compound (2-amino-2-deoxy-D-mannitol, 3) was previously reported as its acetate only for the purpose of analyzing the 2-aminoaldohexoses of bacterial heteroglycans, but no work has been reported on its microbial origin. Since we reported the production of moranoline (1) by Streptomyces sp., we have been studying a more advanced fermentation to improve the productivity of 1 for industrial purposes, and observed a few by-products which interfered with the purification of 1; one of these by-products was revealed to be 1,5-dideoxy-1,5-imino-D-mannitol (2). In the present paper, we describe the isolation of another by-product (3) from the culture broth of Streptomyces lavendulae GC-148, a producing strain of 1 and 2. The strain was cultivated on a rotary shaker and in a jar fermentor with a medium containing soluble starch, soybean meal, yeast extract, NaCl, NaNO3, KCl and MgSO4·7H2O (pH 7.2). The broth filtrate was applied to a Dowex 50Wx2 (H+) column, desorbed with 1 N aqueous ammonia, then to a Diaion SAllA (OH-) column. The physico-chemical properties of the compound were determined: mp 161-163°C, [α]D24 +4.0° (c 1.0, H2O), anal. Calcd. for C6H15NO5: C 39.77, H 8.34, N 7.73; Found: C 39.62, H 8.17, N 7.81, mass spectrum peaks at m/z 182 (M++1, 4.1%) and 183 (M++2, 0.5%), 1H NMR (in D2O) δ 2.98-3.12 (1H, m) and 3.56-3.90 (7H, m), 13C NMR (in D2O) δ 53.84 (CH), 64.05 (CH2), 64.31 (CH2), 70.73 (CH), 71.29 (CH), 71.85 (CH). The compound was identified as 2-amino-2-deoxy-D-mannitol (3) by comparing with synthetic 2-amino-2-deoxy-D-mannitol, which was prepared via sodium borohydride reduction and consecutive N-deacetylation of N-acetyl-D-mannosamine. Furthermore, supplementation of 3 at levels of 0.5% and 1% to the culture medium of S. lavendulae NL-476 (a natural isolate of GC-148) increased the yield of 1 from 3601 µg/ml to 4969 µg/ml and 6181 µg/ml, respectively, suggesting that 3 is closely involved in the biosynthesis of 1.

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