US 4,230,801 A · Production of .alpha.-emulsans

Abstract

Growth of Arthrobacter Sp. ATCC 31012 on fatty acid substrates produces a new class of extracellular microbial protein-associated lipopolysaccharides (the ".alpha.-emulsans"). Deproteinization of .alpha.-emulsans by hot phenol extraction produces the lipopolysaccharide components (the "apo-.alpha.-emulsans") of such emulsans, which components have been shown to be completely N-acylated and partially O-acylated heteropolysaccharides made up of major amounts of D-galactosamine and an aminouronic acid, such apo-.alpha.-emulsans containing at least 5 percent by weight of O-substituted fatty acid esters in which the fatty acids contain from about 10 to about 18 carbon atoms. .alpha.-Emulsans and apo-.alpha.-emulsans, both of which biopolymers are strongly anionic, exhibit a high degree of specificity in the emulsification of hydrocarbon substrates which contain both aliphatic and cyclic components. In addition, these extracellular microbial polysaccharides as well as their O-deacylated and N-deacylated derivatives are adsorbed on and capable of flocculating aluminosilicate ion-exchangers, such as kaolin and bentonite.

Claims23

Dependent claims

A process for producing extracellular microbial lipopolysaccharides which comprises (A) inoculating an aqueous fermentation medium containing a growth-sustaining amount of one or more fatty acid salts with a culture of Acinetobacter Sp. ATCC 31012 or its mutants;

(B) aerobically growing the miocroorganism in such fermentation medium, while adding additional amounts of such fatty acid salt or salts to sustain growth, for a period of time sufficient to produce extracellular microbial protein-associated lipopolysaccharides (herein collectively called ".alpha.-emulsions") in which the lipopolysaccharide components (herein collectively called "apo-.alpha.-emulsans") of such .alpha.-emulsans are N- and O-lipoacylated heteropolysaccharides made up of major amounts of D-galactosamine and an aminouronic acid, such apo-.alpha.-emulsans containing at least 5 percent or above by weight of fatty acid esters in which (1) the fatty acids contain from about 10 to about 18 carbon atoms;
and (2) about 50 percent by weight or more of such fatty acids are composed of 2-hydroxydodecanoic acid and 3-hydroxydodecanoic acid;
and (C) separating substantially all of the microbial cell mass from the emulsan-containing culture medium.

A process for producing extracellular microbial lipopolysaccharides which comprises (A) inoculating an aqueous fermentation medium containing a growth sustaining amount of the assimilable salts of one or more fatty acids with a culture of Acinetobacter Sp. ATCC 31012 or its mutants;

(B) aerobically growing the microorganism in such fermentation medium, while adding additional amounts of such fatty acid salt or salts to sustain growth, aerating the fermentation medium sufficiently to give an oxygen flow rate of about 190 millimoles per liter per hours or higher, and maintaining the pH of the fermentation medium in the range from about 6.2 to about 6.7 and the temperature of the fermentation medium in the range from about 25.degree. C. to about 35.degree. C., for a period of time sufficient to produce extracellular microbial protein-associated lipopolysaccharides (herein collectively called ".alpha.-emulsans") in which the lopopolysaccharide components (herein collectively called "apo-.alpha.-emulsans") are N- and 0-lipoacylated heteropolysaccharides composed of (i) from about 20 to about 35 percent by weight of D-galactosamine;
(ii) from about 30 to about 35 percent by weight of an aminouronic acid;
and (iii) from about 7 to about 19 percent by weight of fatty acid esters in which the fatty acids contain from about 10 to about 18 carbon atoms and are characterized by an average equivalent weight from about 200 to about 230, from about 50 to about 70 percent by weight of such fatty acids being composed of 2-hydroxydodecanoic acid and 3-hydroxydodecanoic acid;
and (C) separating substantially all of the microbial cell mass from the emulsan-containing culture medium.

1 more element

Dependencies are inferred from the claim text.

Production of .alpha.-emulsans

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Production of .alpha.-emulsans

Life of this patent

Abstract

Claims23