GB1067253A - Process for the preparation of enzymes - Google Patents

Process for the preparation of enzymes

Info

Publication number
GB1067253A
GB1067253A GB640365A GB640365A GB1067253A GB 1067253 A GB1067253 A GB 1067253A GB 640365 A GB640365 A GB 640365A GB 640365 A GB640365 A GB 640365A GB 1067253 A GB1067253 A GB 1067253A
Authority
GB
United Kingdom
Prior art keywords
dialysed
aqueous solution
solution
protamine sulphate
chondroitinase
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
Application number
GB640365A
Inventor
Lily Baxendale
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Biorex Laboratories Ltd
Original Assignee
Biorex Laboratories Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Biorex Laboratories Ltd filed Critical Biorex Laboratories Ltd
Priority to GB640365A priority Critical patent/GB1067253A/en
Publication of GB1067253A publication Critical patent/GB1067253A/en
Expired legal-status Critical Current

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)
  • Enzymes And Modification Thereof (AREA)

Abstract

In a process for the production of very pure chondroitinase, cells of Proteus vulgaris (N.C.T.C. 4636) are macerated with a solvent, filtered off and dried, the dried material homogenized in a buffer solution with a pH of about 7.0, incubated at this pH for a short time, the liquid phase separated and dialysed against water, the dialysed, cell-free extract mixed with an aqueous solution of a salt of yeast nucleic acid previously adjusted to a pH of about 7.4, the material which thereby precipitates re-suspended in water and dissolved by the addition of an aqueous solution of an alkali metal hydroxide until the pH is about 7.4, an aqueous solution of protamine sulphate added until the addition of protamine sulphate to a sample clarified by centrifuging no longer gives a precipitate, a slight excess of protamine sulphate then added and the solution as a whole dialysed against water, precipitated material removed, the supernatant liquid adjusted to a pH of about 8.0, cooled, clarified and again dialysed, the resultant clear solution finally being lyophilized. The powder obtained may be adsorbed on calcium phosphate gel and fractionally eluted with sodium acetate, or if it is desired to recover also the chondrosulphatase with which the chondroitinase is contaminated, the two enzymes may be separated chromatographically on DEAE-"Sephadex" (Registered Trade Mark).
GB640365A 1965-02-15 1965-02-15 Process for the preparation of enzymes Expired GB1067253A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
GB640365A GB1067253A (en) 1965-02-15 1965-02-15 Process for the preparation of enzymes

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
GB640365A GB1067253A (en) 1965-02-15 1965-02-15 Process for the preparation of enzymes

Publications (1)

Publication Number Publication Date
GB1067253A true GB1067253A (en) 1967-05-03

Family

ID=9813887

Family Applications (1)

Application Number Title Priority Date Filing Date
GB640365A Expired GB1067253A (en) 1965-02-15 1965-02-15 Process for the preparation of enzymes

Country Status (1)

Country Link
GB (1) GB1067253A (en)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5498536A (en) * 1994-04-22 1996-03-12 American Cyanamid Company Chondroitinase II from Proteus vulgaris
EP0702715A1 (en) * 1993-04-23 1996-03-27 American Cyanamid Company CLONING AND EXPRESSION OF THE CHONDROITINASE I AND II GENES FROM $i(P. VULGARIS)
US5525500A (en) * 1994-04-22 1996-06-11 American Cyanamid Company Chromatographic process for the copurification of chondroitinase I and II proteins from Proteus vulgaris
EP0756636A1 (en) * 1994-04-22 1997-02-05 American Cyanamid Company Chondroitinases i and ii, methods of preparation, and use thereof
US5888798A (en) * 1995-06-07 1999-03-30 American Cyanamid Company Chondroitinase I and chondroitinase II producing mutants of P. vulgaris
WO1999046368A2 (en) * 1998-03-13 1999-09-16 Biomarin Pharmaceuticals Carbohydrate-modifying enzymes for burn and wound debridement and methods for treatment
US6007810A (en) * 1997-05-02 1999-12-28 Seikagaku Corporation Chondroitinase compositions

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0702715A4 (en) * 1993-04-23 2000-01-19 American Cyanamid Co CLONING AND EXPRESSION OF THE CHONDROITINASE I AND II GENES FROM $i(P. VULGARIS)
EP0702715A1 (en) * 1993-04-23 1996-03-27 American Cyanamid Company CLONING AND EXPRESSION OF THE CHONDROITINASE I AND II GENES FROM $i(P. VULGARIS)
US5741692A (en) * 1994-04-22 1998-04-21 American Cyanamid Company Protein vulgaris chondroitinase II
EP0756636A1 (en) * 1994-04-22 1997-02-05 American Cyanamid Company Chondroitinases i and ii, methods of preparation, and use thereof
EP0756636A4 (en) * 1994-04-22 1997-12-17 American Cyanamid Co Chondroitinases i and ii, methods of preparation, and use thereof
US5716617A (en) * 1994-04-22 1998-02-10 American Cyanamid Company Compositions of proteus vulgaris chondroitinase I and chondroitinase II
US5498536A (en) * 1994-04-22 1996-03-12 American Cyanamid Company Chondroitinase II from Proteus vulgaris
US5855883A (en) * 1994-04-22 1999-01-05 American Cyanamid Company Method of disinsertion of vitreous body from neural retina of the eye with Proteus vulgaris chondroitinases I and II
US5525500A (en) * 1994-04-22 1996-06-11 American Cyanamid Company Chromatographic process for the copurification of chondroitinase I and II proteins from Proteus vulgaris
US5888798A (en) * 1995-06-07 1999-03-30 American Cyanamid Company Chondroitinase I and chondroitinase II producing mutants of P. vulgaris
US6007810A (en) * 1997-05-02 1999-12-28 Seikagaku Corporation Chondroitinase compositions
WO1999046368A2 (en) * 1998-03-13 1999-09-16 Biomarin Pharmaceuticals Carbohydrate-modifying enzymes for burn and wound debridement and methods for treatment
WO1999046368A3 (en) * 1998-03-13 2000-08-31 Biomarin Pharmaceuticals Carbohydrate-modifying enzymes for burn and wound debridement and methods for treatment

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