# US 4,400,467 A · Process of using xanthomonas campestris NRRL B-12075 and NRRL B-12074 for making heteropolysaccharide > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 4,400,467 A - Document ID: US4400467A - Status: Granted - Kind: A - Application number: 06/251,831 - Filed: 1981-04-07 - Granted: 1983-08-23 - Earliest claimed: 1980-07-14 - Assignees: Standard Oil Company (Indiana) - Inventors: Keith A. Bauer; Behzad Khosrovi - Web page: https://patentgrep.com/patent/US4400467A ## CPC - C12N 1/205: Bacterial isolates - C12P 19/06: Xanthan, i.e. Xanthomonas-type heteropolysaccharides - C12R 2001/64: Xanthomonas - Y10S 435/813: Continuous fermentation - Y10S 435/91: Xanthomonas - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} - C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS ## Abstract This invention involves the production of heteropolysaccharides such as xanthan gum by the continuous fermentation of suitable nutrient media with new degenerative resistant variants of Xanthomanas campestris. Fermentation is conducted in a nitrogen, sulfur or phosphorous limited nutrient medium containing glucose or a minimal medium plus yeast extract and glutamic acid, in a first fermenter to enhance cultural growth. After the limiting growth element (nitrogen, sulfur or phosphorous) has been essentially consumed, the medium is then transferred to a second fermenter and fermentation continued while adding a suitable carbohydrate which the bacteria convert into the heteropolysaccharide product. The process may be modified by carrying out cultural growth and production of the polysaccharide in the same fermenter. The bacteria employed were cultured from a glucose-mineral salts medium fortified with yeast extract or a glucose-defined minimal medium in which ammonium chloride served as the sole assimilable nitrogen source. ## Claims ### Claim 1 A method for the production of a heteropolysaccharide by means of continuous fermentation which comprises innoculating a liquid carbohydrate minimal medium with a degenerative resistant strain of Xanthomonas campestris selected from the group consisting of Xanthomonas campestris L3 and Xanthomonas campestris L5, having the identifying characteristics of NRRL B-12074 and NRRL B-12075, respectively, cultivating said strain in said medium without discontinuing the fermentation process, withdrawing the results fermented medium therefrom at a rate such that an essentially steady state condition is maintained and recoverying heteropolysaccharide from said fermented medium. ### Claim 2 A novel continuous process for preparing a heteropolysaccharide which comprises continuously cultivating a degenerative resistant microorganism selected from the group consisting of Xanthomonas campestris L3 and Xanthomonas campestris L5, having the identifying characteristics of NRRL B-12074 and NRRL B-12075, respectively, under aerobic conditions in a liquid nutrient medium wherein the growth limiting nutrient is any of the nutrients normally used in said process so that the production of microorganisms is maximized, thereafter continuously feeding the microorganisms thus produced in said medium to a second stage into which a fermentable sugar is fed whereby the formation of heteropolysaccharide in said second stage is maximized. ### Claim 3 A novel continuous process for preparing a heteropolysaccharide which comprises cultivating a degenerative resistant microorganism strain selected from the group consisting of Xanthomonas campestris L3 and Xanthomonas campestris L5 having the identifying characteristics of NRRL B-12704 and NRRL B-12075, respectively, in a liquid carbohydrate nutrient minimal medium containing a complex undefined nitrogen source under aerobic conditions, wherein said medium is continuously fed to a fermentation zone and a culture of one of said strains containing said heteropolysaccharide is continuously withdrawn therefrom. ### Claim 4 (depends on 1) The process of claim 1, wherein said minimal medium contains a complex undefined source of nitrogen. ### Claim 5 (depends on 2) The process of claim 2 wherein the limiting nutrient in said liquid medium is nitrogen. ### Claim 6 (depends on 2) Process of claim 2 wherein the limiting nutrient in said liquid medium in phosphorous. ### Claim 7 (depends on 2) Process of claim 2 wherein the limiting nutrient in said liquid medium is sulfur. ### Claim 8 (depends on 1) Process of claim 1 in which the heteropolysaccharide is xanthan and the carbohydrate is glucose. ### Claim 9 (depends on 1) Process of claim 1 wherein said liquid medium is a glucose-defined mineral salts medium containing a complex undefined nitrogen source. ### Claim 10 (depends on 9) Process of claim 9 wherein said undefined nitrogen source is yeast extract. ### Claim 11 (depends on 9) Process of claim 9 wherein said undefined nitrogen source is yeast autolysate. ### Claim 12 (depends on 1) Process of claim 1 wherein the degenerative resistant strain employed is Xanthomonas campestris B-12074. ### Claim 13 (depends on 1) Process of claim 1 wherein the degenerative resistant strain employed is Xanthomonas campestris B-12075. ### Claim 14 A culture consisting essentially of Xanthomonas campestris L3 NRRL B-12074. ### Claim 15 A culture of a bacterial strain consisting essentially of Xanthomonas campestris NRRL L3 B-12074, said strain being stable and capable of xanthan gum production in continuous fermentation conditions in a nutrient media comprising assimilable sources of carbon, nitrogen, and essential minerals. ### Claim 16 A culture consisting essentially of Xanthomonas campestris L5 NRRL B-12075. ### Claim 17 A culture of a bacterial strain consisting essentially of Xanthomonas campestris L5 NRRL B-12075, said strain being stable and capable of xanthan gum production in continuous fermentation conditions in a nutrient media comprising assimilable sources of carbon, nitrogen, and minerals.