The activity of acis‐9‐fatty acid hydratase produced by aPseudomonas sp. (NRRL B‐3266) isolated from soil was compared with that of another isolate previously reported (NRRL B‐2994). The presence of appropriate fatty acids for at least 4 hr during aerobic growth in yeast extract medium increased subsequent enzyme activity. Such cells anaerobically hydrated severalcis‐9‐alkenoic acids to 10‐hydroxy fatty acids and aerobically formed 10‐keto acids, which were partially degraded to shorter chain keto acids. Melting point, gas chromatography, infrared, mass spectrometry and optical rotatory dispersion data are given. Six fatty acids havingcis‐9‐unsaturation produced hydrated products, but several enoic acids havingtrans‐9‐unsaturation or double bonds in other than the 9 position were inactive as substrates. The (−)‐10‐hydroxypalmitic acid produced from palmitoleic acid is considered to have the D configuration. Yields of 71% crude crystalline product from 15 g of oleic acid and 53% from 11 g of palmitoleic acid were obtained in 5‐liter anaerobic fermentations with NRRL B‐3266. Methyl esters, triolein and oleyl alcohol were not hydrated.

Three new 10‐hydroxy fatty acids, all optically active, have been prepared by the anaerobic microbiological hydration of acis‐9 double bond. Substrates that formed these new hydroxy fatty acids are linoleic, linolenic, and ricinoleic acids. The hydroxyl group has the D configuration and the methyl esters are levorotatory. Infrared, mass spectral, specific rotation and ultraviolet data on these compounds were determined. There was no migration of the unreated double bonds at C12 and C15 in linoleic or linolenic acids. The presence of a double bond in the 10‐hydroxy fatty acids significantly increased the optical rotation of the methyl esters. The hydratase enzyme showed unusual specificity among Δ9 unsaturated acids. While it hydrates methylene interrupted and hydroxy unsaturated acids, it failed to hydrate either 9‐decenoic, 12,13‐epoxy‐ or 12‐keto‐cis‐9‐octadecenoic acids or sterculic acid.

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Aeration of activated sludge with 3 to 4% added methanol for 5 to 7 days yields an odorless, highly viscous (5,000 to 10,000 centipoise), black, pudding-like product containing glycan(s) linked other than a-1-4 or (3-1-3. Backseeding gives maximum thickening in 3 to 4 days. Incomplete acid hydrolysis of the black product gives a 0.27% solution of reducing sugars (75% glucose) which is an 11.4% yield from the added methanol. Backseeding into either centrifuge supernatant or 0.1% yeast extract in tap water gives a light-colored polymer. Viscosity decreases during extended sterile cold storage. A 5% salt addition lowers viscosity one-half. From 6 to 12 colony types appear on plating backseeded media, but none of these isolates is a reliable polymer former.