WIPO logo
Mobile | Deutsch | Español | Français | 日本語 | 한국어 | Português | Русский | 中文 | العربية |
PATENTSCOPE

Search International and National Patent Collections
World Intellectual Property Organization
Search
 
Browse
 
Translate
 
Options
 
News
 
Login
 
Help
 
Machine translation
1. (WO1982000253) IMMOBILIZED AMINOACYLASE ENZYME PREPARATIONS,A PROCESS FOR THE PREPARATION THEREOF,AND A PROCESS FOR THE ISOLATION OF AMINOACYLASE ENZYME FROM THE KIDNEYS OF MAMMALS
Latest bibliographic data on file with the International Bureau   

Pub. No.:    WO/1982/000253    International Application No.:    PCT/HU1981/000008
Publication Date: 04.02.1982 International Filing Date: 02.03.1981
IPC:
C12N 11/08 (2006.01), C12N 9/80 (2006.01), C12P 13/04 (2006.01)
Applicants: BOROS, László; (For US Only).
DARÓCZI, Iván; (For US Only).
HUBER, Irén; (For US Only).
IVONY, Józsefné; (For US Only).
KISS, Jánosné; (For US Only).
SZAJÁNI, Béla; (For US Only).
REANAL FINOMVEGYSZERGYÁR (For All Designated States Except US)
Inventors: BOROS, László; .
DARÓCZI, Iván; .
HUBER, Irén; .
IVONY, Józsefné; .
KISS, Jánosné; .
SZAJÁNI, Béla;
Priority Data:
Title (EN) IMMOBILIZED AMINOACYLASE ENZYME PREPARATIONS,A PROCESS FOR THE PREPARATION THEREOF,AND A PROCESS FOR THE ISOLATION OF AMINOACYLASE ENZYME FROM THE KIDNEYS OF MAMMALS
(FR) PREPARATIONS D'AMINO-ACYLASE IMMOBILISE, PROCEDE DE PREPARATION DE CELLE-CI, ET PROCEDE D'ISOLATION DE L'AMINO-ACYLASE A PARTIR DES REINS DE MAMMIFERES
Abstract: front page image
(EN)An improved method for the isolation of aminoacylase enzyme from mammal kidneys. According to the invention an aqueous kidney extract, prepared in a manner known per se, is subjected to heat treatment at 60 to 80 C for 5 to 15 minutes, the heat treated mixture is centrifuged, 200 to 300 g/litre of a salt of the general formula KxA (wherein K stands for a monovalent cation, A is the anion of a mineral acid and x is equal to the valence of the anion) are added to the supernatant, the resulting suspension is centrifuged, the separated precipitate is dissolved in water, the solution is dialyzed against water, and, if desired, the dialyzed solution or a solution of the enzyme, separated from the dialyzate, in a buffer of pH 6.5 to 8.5 is subjected to anion exchange chromatography, finally, if desired, the active fractions obtained in the chromatographic step or a solution of the enzyme, precipitated from the active fractions with a salt of the general formula KxA, in a buffer of pH 6.0 to 8.0 is subjected to gel chromatography on a gel filter wherein the upper limit of the fractionation molecular weight is higher than 70,000, and the enzyme is precipitated from the active fractions with a salt of the general formula KxA as described above. The invention also relates to an immobilized aminoacylase enzyme consisting of a partially hydrolyzed A krilex P type acrylamide-N,N'-methylene-bis(acrylamide) copolymer and an aminoacylase enzyme attached to said copolymer by covalent bonds This immobilized enzyme preparation is prepared according to the invention so that a partially hydrolyzed Akrilex P type copolymer is treated with a carbodiimide compound which is soluble in water or soluble in an organic solvent at a temperature below 0 C, a solution of the aminoacylase with a pH of 6.5 to 8.5 is applied onto the resulting activated support, the resulting product is washed and then dried, if desired. By the improved method of the invention aminoacylase can be separated from mammal kidneys far more easily than before. The immobilized enzyme preparations according to the invention are of extremely high specific activity.
(FR)Procede ameliore permettant l'isolation de l'amino-acylase a partir des reins de mammiferes. Selon l'invention un extrait aqueux de reins, prepare selon un procede connu, est soumis a un traitement calorifique a une temperature allant de 60 a 80 C pendant 5 a 15 minutes, le melange traite par la chaleur est centrifuge, 200 a 300g/litres d'un sel correspondant a la formule generale KxA (dans laquelle K represente un cation monovalent, A represente l'anion d'un acide mineral et x est egal a la valence de l'anion) sont ajoutes au surnageant, la suspension obtenue est centrifugee, le precipite separe est dissout dans de l'eau, la solution est dialysee contre l'eau et, facultativement, la solution dialysee ou une solution de l'enzyme, separee du dialysat, dans un tampon d'un pH allant de 6,5 a 8,5 est soumise a une chromatographie a echange anionique, et finalement, facultativement, les fractions actives obtenues dans l'etape chromatographique ou une solution de l'enzyme, precipitee des fractions actives au moyen d'un sel correspondant a la formule generale KxA, dans un tampon d'un pH allant de 6,0 a 8,0 sont soumises a une chromatographie sur gel sur un filtre dans laquelle la limite superieure du poids moleculaire de fractionnement est superieure a 70000, et l'enzyme est precipite des fractions actives au moyen d'un sel correspondant a la formule generale KxA deja decrite. L'invention se refere aussi a une amino-acylase immobilisee se composant d'un copolymere partiellement hydrolyse d'une acrylamide-M,M'-methylene-bis(acrylamide) du type Acrylex P et d'une amino-acylase liee a ce copolymere par des liaisons covalentes. Cette preparation d'enzyme immobilise est preparee selon l'invention de maniere qu'un copolymere du type Acrylex P partiellement hydrolyse soit traite avec un compose carbodiimide qui est soluble dans l'eau ou dans un solvant organique a une temperature inferieure a 0 C, une solution de l'amino-acylase presentant un pH allant de 6,5 a 8,5 est applique sur le support active resultant, le produit resultant est lave et ensuite seche facultativement. Par ce procede ameliore l'amino-acylase peut etre separee des reins de mammiferes beaucoup plus facilement qu'auparavant. Les preparations d'enzyme immobilise selon l'invention presentent une activite specifique extremement elevee.
Designated States:
Publication Language: English (EN)
Filing Language: English (EN)