Method for preparing cytidine diphosphate choline

Abstract

Cytidine diphosphate choline is prepared by culturing an aqueous reaction liquor containing (a) at least one of the class consisting of choline or phosphoryl choline; (b) at least one of the class consisting of cytidine, cytidine-5'-monophosphate, cytidine-5'-diphosphate or cytidine-5'triphosphate; (c) enzyme systems of bacteria, yeasts or molds; (d) phosphate ion and (e) at least one of the class consisting of magnesium ion and manganese ion and recovering cytidine diphosphate choline from said culturing liquor.

What is claimed is: .[.1. A method for preparing cytidine diphosphate choline, which comprises culturing an aqueous reaction liquor containing (a) at least one of the class consisting of choline of phosphoryl choline; (b) at least one of the class consisting of cytidine, cytidine-5'-monophosphate, cytidine-5'-diphosphate and cytidine-5'-triphosphate; (c) cells of bacteria yeasts or molds that are in a form which liberate their enzyme systems into the aqueous reaction liquor under the culturing conditions; (d) phosphate ion (e) at least one of the class consisting of magnesium ion and manganese ion (f) at least one fermentable sugar and recovering cytidine diphosphate choline from 
     
        said culturing liquor..]. 2. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein said culturing is carried out at a temperature of from 
     
     
        about 20° C. to 55° C. 3. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein the pH of said aqueous reaction liquor 
     
     
        is from about 5.0 to 9.0. 4. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein (a) comprises choline and (b) comprises 
     
     
        cytidine-5'-monophosphate. 5. A method according to claim .[.1.]. .Iadd.14 
     
     
        .Iaddend.wherein (a) comprises choline and (b) comprises cytidine. 6. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein (a) comprises 
     
     
        choline and (b) comprises cytidine-5'-diphosphate. 7. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein (a) comprises choline and (b) 
     
     
        comprises cytidine-5'-triphosphate. 8. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein said phosphate ion is supplied by a mixture of 
     
     
        monopotassium phosphate and dipotassium phosphate. 9. A method according 
     
     
        to claim .[.1.]. .Iadd.14 .Iaddend.(e) comprises magnesium sulfate. 10. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein said 
     
     
        fermentable sugar comprises glucose. 11. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein recovery of said cytidine diphosphate choline is effected by passing a filtrate containing the cytidine diphosphate choline obtained from the aqueous reaction liquor through an 
     
     
        ion exchange resin column. 12. A method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein (c) is at least one member of the class consisting of: Brevibacterium ammoniagenes ATCC 6872   Escherichia coli ATCC 21148   Serratia mercescens ATCC 2123   Aerobacter aerogenes ATCC 21217   Corynebacterium glutamicum ATCC 13287   Arthrobacter simplex ATCC 15799   Micrococcus sodonensis ATCC 11880   Pseudomonas fluorescens ATCC 21256   Bacillus subtilis ATCC 15512   Nocardia globerula ATCC 21022   Streptomyces ambofaciens ATCC 15154   Penicillium chrysogenum ATCC 15241   Aspergillus terreus ATCC 1012   Aspergillus flavus ATCC 13698   Gibberella fujikuroi ATCC 20136   Rhizopus delemar ATCC 20134   Mucor javanicus ATCC 15242   .[.saccharomyces cerevisiae ATCC 15248   Saccharomyces lactis ATCC 12425   Torulopsis sphaerica ATCC 8549   Zygosaccharomyces major (synonym Saccharomyces rouxii) ATCC 15249   Kloeckera africana ATCC 16512   Candida guilliermondii ATCC 9058.]. 13. a method according to claim .[.1.]. .Iadd.14 .Iaddend.wherein said fermentable sugar is present in a concentration from about 5 g./l. to 100 g./l. of said reaction liquor. .Iadd. 14. A method for preparing cytidine diphosphate choline which comprises culturing an aqueous reaction liquor containing (a) at least one of the class consisting of choline and phosphoryl choline; (b) at least one of the class consisting of cytidine, cytidine-5' -monophosphate, cytidine- 5' -diphosphate and cytidine- 5' -triphosphate; (c) cells of bacteria or molds that are in a form which liberate their enzyme systems into the aqueous reaction liquor under the culturing conditions; (d) phosphate ion (e) at least one of the class consisting of magnesium ion and manganese ion; (f) at least one fermentable sugar and recovering cytidine diphosphate choline from said culture liquor.