GB1060513A - Process for the preparation of glucuronoglycosaminoglycan hyaluronate lyase - Google Patents
Process for the preparation of glucuronoglycosaminoglycan hyaluronate lyaseInfo
- Publication number
- GB1060513A GB1060513A GB640465A GB640465A GB1060513A GB 1060513 A GB1060513 A GB 1060513A GB 640465 A GB640465 A GB 640465A GB 640465 A GB640465 A GB 640465A GB 1060513 A GB1060513 A GB 1060513A
- Authority
- GB
- United Kingdom
- Prior art keywords
- precipitate
- solution
- treating
- mls
- adjusting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
Hyaluronidase (glucuronoglycosaminoglycan hyaluronate lyase) is prepared from animal organs or products thereof containing the said enzyme, by: i) macerating with aqueous acetic acid containing 2.5 to 10% by volume hydrochloric acid, and centrifuging off the solids, ii) treating the supernatant with 21 g. ammonium sulphate per 200 mls. solution and removing the precipitate, iii) treating the resulting supernatant with 56.5 g. ammonium sulphate per 200 mls. solution, collecting the precipitate, and dissolving it in the minimum amount of water, iv) ultra-filtering this solution to 1/10 of its original volume, and dialysing the concentrae against water, v) treating the dialysate with two volumes of chloroform, separating the aqueous layer, centrifuging off any precipitate, and adjusting to pH 2.3, and standing for 1 to 15 minutes, then adjusting to pH 4.8, and removing the precipitate, vi) treating the clarified solution with 10 g. ammonium sulphate per 100 mls, and collecting the precipitae, vii) dissolving the precipitate in water and dialysing against physiological saline, viii) chromatographing the dialysate through a column of cross-linked sulphonated polystyrene ion exchange resin and eluting with pH 1.5 KCl/HCl buffer, ix) adjusting the pH of the elurate to 6.0 and ultra-filtering to 1/20 of its original volume and x) lyophilising the concentrated solution, if required. The product may be further purified by preparative electrophoresis on starch gel or polyacrylamide gel. Suitable starting materials are bulls' testicles or liver lysozymes. The product is said to have an activity of at least 21,500 International units and to be almost free from contaminating enzymes.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB640465A GB1060513A (en) | 1965-02-15 | 1965-02-15 | Process for the preparation of glucuronoglycosaminoglycan hyaluronate lyase |
ES0323078A ES323078A1 (en) | 1965-02-15 | 1966-02-15 | Procedure for the preparation of enzymes of glucuronoglicosaminoglican hialuronato liasa. (Machine-translation by Google Translate, not legally binding) |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB640465A GB1060513A (en) | 1965-02-15 | 1965-02-15 | Process for the preparation of glucuronoglycosaminoglycan hyaluronate lyase |
Publications (1)
Publication Number | Publication Date |
---|---|
GB1060513A true GB1060513A (en) | 1967-03-01 |
Family
ID=9813914
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
GB640465A Expired GB1060513A (en) | 1965-02-15 | 1965-02-15 | Process for the preparation of glucuronoglycosaminoglycan hyaluronate lyase |
Country Status (1)
Country | Link |
---|---|
GB (1) | GB1060513A (en) |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4152212A (en) * | 1976-02-25 | 1979-05-01 | Biorex Laboratories Limited | Process for the purification of glucuronoglycosaminoglycan hyaluronate lyase |
FR2495634A1 (en) * | 1980-12-08 | 1982-06-11 | Biorex Laboratories Ltd | ENZYME NEW AND USEFUL IN THERAPEUTICS |
EP0355831A2 (en) * | 1988-08-24 | 1990-02-28 | Seikagaku Kogyo Co., Ltd. | Purification of glycosaminoglycan degrading enzymes |
US6689349B1 (en) | 1998-12-23 | 2004-02-10 | Esparma Gmbh | Skin protection agents containing a fragment mixture produced from hyaluronic acid by hydrolysis |
US6719986B1 (en) | 1998-12-23 | 2004-04-13 | Esparma Gmbh | Hyaluronate lyase used for promoting penetration in topical agents |
CN109722424A (en) * | 2019-01-25 | 2019-05-07 | 山东大学 | A method of the separate lactose enzyme from infant faeces |
-
1965
- 1965-02-15 GB GB640465A patent/GB1060513A/en not_active Expired
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4152212A (en) * | 1976-02-25 | 1979-05-01 | Biorex Laboratories Limited | Process for the purification of glucuronoglycosaminoglycan hyaluronate lyase |
FR2495634A1 (en) * | 1980-12-08 | 1982-06-11 | Biorex Laboratories Ltd | ENZYME NEW AND USEFUL IN THERAPEUTICS |
EP0355831A2 (en) * | 1988-08-24 | 1990-02-28 | Seikagaku Kogyo Co., Ltd. | Purification of glycosaminoglycan degrading enzymes |
EP0355831A3 (en) * | 1988-08-24 | 1991-08-28 | Seikagaku Kogyo Co., Ltd. | Purification of glycosaminoglycan degrading enzymes |
US5198355A (en) * | 1988-08-24 | 1993-03-30 | Seikagaku Kogyo Co., Ltd. | Purification of glycosaminoglycan degrading enzymes with a sulfated polysaccharide |
US6689349B1 (en) | 1998-12-23 | 2004-02-10 | Esparma Gmbh | Skin protection agents containing a fragment mixture produced from hyaluronic acid by hydrolysis |
US6719986B1 (en) | 1998-12-23 | 2004-04-13 | Esparma Gmbh | Hyaluronate lyase used for promoting penetration in topical agents |
CN109722424A (en) * | 2019-01-25 | 2019-05-07 | 山东大学 | A method of the separate lactose enzyme from infant faeces |
CN109722424B (en) * | 2019-01-25 | 2022-08-05 | 山东大学 | Method for separating lactase from infant excrement |
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