# US 5,703,040 A · Broad spectrum antibiotic peptide > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 5,703,040 A - Document ID: US5703040A - Status: Granted - Kind: A - Application number: 08/561,935 - Filed: 1995-11-22 - Granted: 1997-12-30 - Earliest claimed: 1995-11-22 - Assignees: Kansas State University Research Foundation - Inventors: John J. Iandolo; Scott Crupper - Web page: https://patentgrep.com/patent/US5703040A ## CPC - C07K 14/31: from Staphylococcus (G) - A01N 63/50: Isolated enzymes; Isolated proteins (peptides A01N37/46) - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - Y10S 530/825: Bacteria - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS ## Abstract Improved antimicrobial proteinaceous substances produced by Staphylococcus aureus KSI1829 (e.g., Bac1829) and methods of inhibiting microbial growth using such substances are disclosed. ## Claims ### Claim 1 An at least partially purified antimicrobial proteinaceous substance produced by Staphyloccocus aureus having a molecular weight of from about 6 to 7 kilodaltons, wherein said substance at a concentration of 640 antimicrobial units/ml in an aqueous solution retains essentially all of its antimicrobial activity after one or more of the following treatments: (1) heat treatment at 95.degree. C. for 15 minutes, (2) treatment with 6M urea or 10 mM dithiothreitol at room temperature for 1 hour, and (3) treatment with deoxyribonuclease, ribonuclease, or lysostaphin at a concentration of 1 mg/ml at room temperature for 1 hour. ### Claim 2 (depends on 1) The substance of claim 1, wherein said Staphylococcus is Staphylococcus aureus KSI1829. ### Claim 3 (depends on 1) The substance of claim 1, wherein said Staphylococcus is grown in a cell culture. ### Claim 4 (depends on 1) The substance of claim 1, wherein said molecular weight is about 6.4 kilodaltons. ### Claim 5 (depends on 1) The substance of claim 1, wherein said substance is a protein. ### Claim 6 (depends on 1) The substance of claim 1, wherein said substance is Bac1829. ### Claim 7 (depends on 1) The substance of claim 1, wherein said substance at a concentration of 640 AU/ml in an aqueous solution retains its antimicrobial activity after being heated at 95.degree. C. for 15 minutes. ### Claim 8 (depends on 1) The substance of claim 1, wherein said substance at a concentration of 640 AU/ml in an aqueous solution retains its antimicrobial activity after being treated at room temperature for 1 hour with urea at a concentration of 6M. ### Claim 9 (depends on 1) The substance of claim 1, wherein said substance at a concentration of 640 AU/ml in an aqueous solution retains its antimicrobial activity after being treated at room temperature for 1 hour with dithiothreitol at a concentration of 10 mM. ### Claim 10 (depends on 1) The substance of claim 1, wherein said substance at a concentration of 640 AU/ml in an aqueous solution retains its antimicrobial activity after being treated at room temperature for 1 hour with an enzyme at a concentration of 1 mg/ml, said enzyme being selected from the group consisting of deoxyribonuclease, ribonuclease, and lysostaphin. ### Claim 11 (depends on 1) The substance of claim 1, wherein said substance at a concentration of 640 AU/ml in an aqueous solution loses its antimicrobial activity after being treated at room temperature for 1 hour with an enzyme at a concentration of 1 mg/ml, said enzyme being selected from the group consisting of proteinase K and trypsin. ### Claim 12 (depends on 9) The substance of claim 9, wherein said substance at a concentration of at least 1280 AU/ml has antimicrobial activity against a microorganism selected from the group consisting of Staphyloccocus aureus 8325-4r, Streptococcus suis, Corynebacterium renale, Corynebacterium pseudotuberculosis Whetten 1, Corynebacterium diptheriae, Haemophilus parasuis, Bordetella pertussis, Bordetella brochoseptica, Moraxella bovis, and Pasteurella multocida. ### Claim 13 (depends on 12) The substance of claim 12, wherein said antimicrobial activity inhibits the growth of said microorganism. ### Claim 14 (depends on 13) The substance of claim 13, wherein said antimicrobial activity prevents the growth of said microorganism. ### Claim 15 A method of inhibiting microbial growth in an environment capable of sustaining said growth comprising administering to said environment a microbial growth-inhibiting amount of an at least partially purified antimicrobial proteinaceous substance produced by Staphylococcus aureus having a molecular weight of from about 6 to 7 kilodaltons, wherein said substance at a concentration of 640 antimicrobial units/ml in an aqueous solution retains essentially all of its antimicrobial activity after one or more of the following treatment: (1) heat treatment at 95.degree. C. for 15 minutes, (2) treatment with 6M urea or 10 mM dithiothreitol at room temperature for 1 hour, and (3) treatment with deoxyribonuclease, ribonuclease, or lysostaphin at a concentration of 1 mg/ml at room temperature for 1 hour. ### Claim 16 (depends on 15) The method of claim 15, wherein said method prevents said microbial growth. ### Claim 17 (depends on 15) The method of claim 15, wherein said Staphylococcus is Staphylococcus aureus KSI1829. ### Claim 18 (depends on 15) The method of claim 15, wherein said Staphylococcus is grown in a cell culture. ### Claim 19 (depends on 15) The method of claim 15, wherein said molecular weight is about 6.4 kilodaltons. ### Claim 20 (depends on 15) The method of claim 15, wherein said substance is a protein. ### Claim 21 (depends on 15) The method of claim 15, wherein said substance is Bac1829. ### Claim 22 (depends on 15) The method of claim 15, wherein said substance at a concentration of 640 AU/ml in an aqueous solution retains its antimicrobial activity after being heated at 95.degree. C. for up to 15 minutes. ### Claim 23 (depends on 15) The method of claim 15, wherein said substance at a concentration of 640 AU/ml in an aqueous solution retains its antimicrobial activity after being treated at room temperature for 1 hour with urea at a concentration of 6M. ### Claim 24 (depends on 15) The method of claim 15, wherein said substance at a concentration of 640 AU/ml in an aqueous solution retains its antimicrobial activity after being treated at room temperature for 1 hour with dithiothreitol at a concentration of 10 mM. ### Claim 25 (depends on 15) The method of claim 15, wherein said substance at a concentration of 640 AU/ml in an aqueous solution retains its antimicrobial activity after being treated at room temperature for 1 hour with an enzyme at a concentration of 1 mg/ml, said enzyme being selected from the group consisting of deoxyribonuclease, ribonuclease, and lysostaphin. ### Claim 26 (depends on 15) The method of claim 15, wherein said substance at a concentration of 640 AU/ml in an aqueous solution loses its antimicrobial activity after being treated at room temperature for 1 hour with an enzyme at a concentration of 1 mg/ml, said enzyme being selected from the group consisting of proteinase K and trypsin. ### Claim 27 (depends on 1) The method of claim 1, wherein said substance at a concentration of at least 1280 AU/ml has antimicrobial activity against a microorganism selected from the group consisting of Staphyloccocus aureus 8325-4r, Streptococcus suis, Corynebacterium renale, Corynebacterium pseudotuberculosis Whetten 1, Corynebacterium diptheriae, Haemophilus parasuis, Bordetella pertussis, Bordetella brochoseptica, Moraxella bovis, and Pasteurella multocida. ### Claim 28 (depends on 27) The method of claim 27, wherein said antimicrobial activity inhibits the growth of said microorganism. ### Claim 29 (depends on 28) The method of claim 28, wherein said antimicrobial activity prevents the growth of said microorganism. ### Claim 30 (depends on 1) The substance of claim 1, said substance being a purified peptide. ### Claim 31 (depends on 15) The method of claim 15, said substance being a purified peptide.