Submitted to: Applied and Environmental Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/1/2003
Publication Date: 7/15/2003
Citation: HOU, C.T., HOSOKAWA, M., WEISLEDER, D. PRODUCTION OF NOVEL TETRAHYDROXYFURANYL FATTY ACIDS FROM A-LINOLENIC ACID BY CLAVIBACTER SP. ALA2. APPLIED AND ENVIRONMENTAL MICROBIOLOGY. 2003. V. 69. P. 3868-3873. Interpretive Summary: Previously, we discovered a new microbial strain ALA2 that produced seven new hydroxy fatty acid products from linoleic acid (a component of soybean oil). Hydroxy fatty acids are useful as starting materials for the synthesis of specialty chemicals, special military nylon, plastisizers, coatings and physiologically active agents. Recently, we expanded our substrate to other polyunsaturated fatty acids, such as alpha-linolenic acid (also a component of soybean oil). We found that strain ALA2 converted alpha-linlenic acid to two novel hydroxy furanyl (five membered ring) fatty acids. Furanyl fatty acids are known anticancer drugs. The new products have great potential to be used as bioactive agents and raw materials for synthesis of specialty chemicals. Application of these products either as agents for anti-plant pathogenic fungi or raw material for synthesis of specialty chemicals will benefit U.S. farmers.
Technical Abstract: Previously, we reported that a newly isolated microbial culture, Clavibacter sp. ALA2 produced trihydroxy unsaturated fatty acids, diepxoy bicyclic fatty acids, and tetrahydroxyfuranyl fatty acids (THFAs) from linoleic acid (12, 16-18). Now, we found that Clavibacter sp. ALA2 produced novel THFAs: 13,16-dihydroxy-12, 15-epoxy-9(Z)-octadecenoic acid (13,16-dihydroxy-THFA) and 7,13,16-trihydroxy-12, 15-epoxy-9(Z)-octadecenoic acid (7,13,16-trihydroxy-THFA) from a-linolenic acid. The chemical structures of these products were determined by GC/MS and proton and 13C NMR analyses. The optimum incubation temperature was 30OC for production of both hydroxy-THFAs. 13,16-Dihydroxy-THFA was detected after 2 days of incubation and reached 45 mg/50ml after 7 days of incubation. Whereas 7,13,16-trihydroxy-THFA wasn¿t detected after 2 days of incubation but reached 9 mg/50ml after 7 days of incubation. Total yield of both 13,16-dihydroxy-THFA and 7,13,16-trihydroxy-THFA reached 67% (w/w) after 7days of incubation at 30OC, 200rpm. In our previous studies, we reported that Clavibacter sp. ALA2 oxidized C-7, C-12, C-13, C-16 and C-17 positions of linoleic acid (n-6) into hydroxy groups. In this case, the bond between C-16 and C-17 carbons is saturated. With a-linolenic acid (n-3), however, the bond between C-16 and C-17 carbons is unsaturated. It seems that enzymes of strain ALA2 oxidized the C-12/C-13 and C-16/C-17 double bonds into dihydroxy groups first, and then converted them to hydroxy-THFAs.