US006066484A United States Patent (19) 11 Patent Number: 6,066,484 Hatanaka et al. (45) Date of Patent: May 23, 2000

54) PURINE NUCLEOSIDASE Takagi et al., “Purification and Properties of a Bacterial Riboside ” (XP-002104771, p. 77–86) from The 75 Inventors: Haruyo Hatanaka, Kyoto; Toshihiko Journal of Biological Chemistry, vol. 225, 1995. Ashikari, Takatsuki; Jun Ogawa; Sakayu Shimizu, both of Kyoto, all of Tofler et al., “A 13-Year Follow-Up of Social Drinkers”, Japan (XP-002104772, p. 479–481) from The Medical Journal of Australia, vol. 2, Oct. 31, 1981. 73 Assignee: Suntory Limited, Osaka, Japan Thompson et al., “The Effect of Inositol and Nucleic Acid Bases on the Fermentation Rate of an Ale Yeast,” A.S.B.C. 21 Appl. No.: 08/912,560 Proceedings, 1973, p. 137–141. 22 Filed: Aug. 18, 1997 Yoshino et al., “ Nucleosidase from Azotobacter 30 Foreign Application Priority Data Vinelandii, Archives of Microbiology, vol. 119, 1978, p. 59-64. Aug. 16, 1996 JP Japan ...... 8-216421 Parkin et al., “Nucleoside Hydrolase from Crithidia Fas (51) Int. Cl." ...... C12N 9/16; C12C 1/00 ciculata.” The Journal of Biochemistry, 1991, p. 52 U.S. Cl...... 435/196; 435/93 20658-20665. 58 Field of Search ...... 435/196, 93 56) References Cited Primary Examiner Karen Cochrane Carlson ASSistant Examiner-Devesh Srivastava U.S. PATENT DOCUMENTS Attorney, Agent, or Firm-Burns, Doane, Swecker & 5,200,313 4/1993 Carrico ...... 435/6 Mathis, L.L.P. FOREIGN PATENT DOCUMENTS 57 ABSTRACT 753 572 1/1997 European Pat. Off.. Novel purine nucleosidase derived from Ochrobactrum anthropi microorganisms, a gene System coding therefor, OTHER PUBLICATIONS uses therefor and particularly its use in the production of Leung et al. Structure and regulation of the AMP nucleosi beer. dase gene (amn) from Escherichia coli. Biochemistry 28: 8726–8733, 1989. 2 Claims, 11 Drawing Sheets

GUANOSINE GUANINE

ADENOSINE e S. 3OO 2 ADENINE

a. S 200

2y CD 25 100 O

ENZYME CONTROL TREATMENT U.S. Patent May 23, 2000 Sheet 1 of 11 6,066,484 NaCl CONCENTRATION (M) ---- d CO CO Nr on ( - O O O O

2NOILOW}},{ 0LL09L09L07L08LOZLOLL00L06080/,070Z0?0 ‘ONNOIJLOWH){

000€ 009Z 000Z 009L 009 <[ CO UT) ÇN CO CD GHAIJ,0W000CL E