270 Patent Survey

culture of the above microorganism has been deposited with the American Type Culture Collection (ATCC) on May 15. 1990.

The polysaccharide from Erwina sp. is characterized by a molecular weight of approximately 2.5 x 106. Its main build- ing blocks are mannose and galactose, which is a component of lactose in whey. Erwina sp. produces the novel galacto- mannan polysaccharide when cultured in the presence of lactose, a component of whey, in addition to other carbohydrates.

The microorganism was isolated from a soil sample of a Wisconsin farm field near Mineral Point, WI, which had been regularly treated with whey.

The microorganism grows rapidly on agar plates containing a preferred medium. which is prepared by adding 1.6% agar to the preferred liquid medium listed in Table 1.

Tahle 1.

Preferred Medium for Fermentation of Erw-inia sp. ATCC 5.5046

Component Concentration

(g/O

Lactose.H,O (NH&SO, KH?PO, K,HPO, MgS04 7H20 CaC1, .21110 FeSO, .7H,O CoClz . 6H20 ZnSOj 7H20 &SO., . 5HZ0 MnSO, .H,O NazMoO, 211z0 Yeast extract NaOH

45.0 1.46 I.8 3.6 0.6 0.04 0~0019 0.00 1 0.00 1 0.00 1 0~001 0,001 1.8

to pH 7 (approx. 0.18 gl)

The novel galactomannan polysaccha- ride recovered by the fermentation pro- cess (which involves ccntrifugation, precipitation with a short-chain alcohol and, optionally, a salt solution, ultrafiltra- tion or ion exchange or dialysis, and vacuum drying or lyophilization) has use- ful properties, primarily for modification of the rheology of aqueous solutions.

Culture concentrates for direct vat set dairy products production

(US patent application 5 128 20, Sanofi Bio Ingredients, Inc., Waukesha, Wisconsin, USA)

Mathison has invented a process for pre- paring concentrated bacterial cultures for direct vat inoculation of milk batches in

which harmless lactic acid-producing bacteria are cultured in an aqueous medium, including fermentation nutrients, to obtain a fermented aqueous culture of the bacteria cells (e.g. of the streprococcus lactic type) together with residual nutrients including casein. It con- sists of dissolving in the culture medium, prior to the recovery and concentration of the cells by centrifugation, a combina- tion of 0.05% to 0.20% (based on the weight of the culture) of a water-soluble food-acceptable poly-phosphate salt, selected from the class consisting of tri- polyphosphate salt and hexametaphos- phate salt and 0.25% to 0.50% (based on the weight of the culture) of a water-solu- ble food-acceptable citrate salt.

It is important that the polyphosphate salt/citrate salt combination dissolves in the maximum amount of solids in the medium before centrifugation. For this purpose the polyphosphate Fait/citrate sail combination can be added prior to, or after, the fermentation. Preferably, the whole combination is added prior to the fermentation since it has been observed that the low levels in polyphosphate salt/ citrate salt required do not interfere in the normal fermentation process and do not inhibit the growth of the bacteria. In addi- tion, the polyphosphate salt, in sufficient amounts, could control bacteriophage during fermentation.

The polyphosphate salt/citrate salt combination used in the process, when added prior to fermentation, provides over a YO%-reduction in solids using several concentrations of polyphosphate salt and citrate salt.

Under such conditions, the culture growth appears to be slightly enhanced both in activity and cell counts. This could be an added benefit ffom the pre- growth casein solubilization.

The hexametaphosphate salt, i.e. com- mercial mixture of polyphosphate salts containing 4 to 22 phosphate groups per molecule, can be cited as the polyphos- phate salt particularly useful in the pro- ccss.

The production of highly unsaturated The Netherlands Patent Oflice, Rijswijk, fatty acid The Netherlands:

(Japanese patent application 63 53641 and US patent 5 128 250, Suntory Ltd, Osaka, Japan)

Akimoto and co-researchers have dis- covered a process for the production of a highly unsaturated fatty acid having an odd number of carbon atoms by culturing a microorganism belonging to the genus Mortierella and capable of producing the fatty acid; typically represented by 8,11,14-nonadecatrienoic acid, produced

The Patent Information Department of TNO, Rijswijk The Netherlands; The International Patent Research Ofire IRPO, PO Box 16260, 2500 BG, The Hague, The Netherlands; UK Patents Ojj?ce, Concept House, Car- diff Road, Newport, Gwent h’P9 IRH, UK; Univento, PO Box IhoS6, The Hague, The Netherlands; USA Patent Office, Box 4, Patent and Trademark Ojice, US Department of Commerce, Washington DC20231, USA.

by culturing a microorganism capable of producing arachidonic acid.

One example of the process is given in the following description.

Two ml of a medium containing 2.0% glucose and 1% yeast extract (pH 6.0) was put into Erlenmeyer flasks, and auto- claved at 120°C for 20 min. Mortierella alpina CBS 219.35, and Mortierella hy- grophila IF0 5941 were separately ino- culated to the same medium, and cultured on a reciprocating shaker at 110 rpm, for seven days at 28°C. After culturing, each culture broth was filtered to obtain cul- tured cells, which were then washed with water and dried in a centrifugal evapora- tor at 60°C for 2 h. To the dried cells 2 ml of methylene chloride and 2 ml of 10% hydrochloric acid in absolute methanol were added, and the whole was incubated at 50°C for 3 h to ester@ fatty acids pro- duccd. To this mixture 4 ml of n-hexane and 1 ml of water were added to extract the fatty acid methyl esters. The extrac- tion was carried out twice, and the extractants were combined and evapo- rated in a centrifugal evaporatory at 40°C for 1 h to eliminate the solvent. The fatty acid methyl esters thus obtained were analyzed by gas chromatography. As a result, MortiereNa alpina CBS 219.35 and Mortierella hydrophila IF0 5941 pro- duced 5,8,11,14-nonadecatetraenoic acid in amounts of 3 mg/l culture broth and 1 mg/l culture broth, respectively.

5,&l 1,14-nonadecatetraenoic acid was isolated by separating the fatty acid methyl esters obtained, as described above, using high performance liquid chromatography on a reverse column (5ClS) with acetonitrile/-water (85: 15) as the eluate. The 5,8,11,14-nonadeca- tetetraenoic acid obtained was confirmed by mass spectrometry and NMR spec- trometry.

Copies of the references cited can be obtained from the following organizations, or at the Patent Offices of the respective countries.

European Patent Office, Rijswijk The Netherlands;