公开(公告)号：US3634196A

公开(公告)日：1972-01-11

申请号：US3634196D

申请日：1968-12-26

Applicant: BAYER AG

Inventor： WAGNER OTTO ,  BAUER KLAUS ,  KAUFMANN WILFRIED ,  RAUENBUSCH ERICH ,  ARENS ALFRED ,  IRION ECKARD

IPC: C12N9/82 ,  C07G7/02

CPC classification number: C12N9/82 ,  Y10S435/815 ,  Y10S435/849

Abstract: Crude L-asparaginase is freed of pyrogens by contacting solutions of the same containing weak buffers with diethylaminoethyl dextran gels. The process can be carried out according to batch techniques or in a continuous manner by using a chromatographic column.

Abstract translation: 通过将含有弱缓冲液的溶液与二乙基氨基乙基葡聚糖凝胶接触来解决热L-天冬酰胺酶，释放热原。 该方法可以根据间歇技术或通过使用色谱柱以连续方式进行。

公开(公告)号：US3650902A

公开(公告)日：1972-03-21

申请号：US3650902D

申请日：1969-05-28

Applicant: BAYER AG

Inventor： WAGNER OTTO ,  BAUER KLAUS ,  KAUFMANN WILFRIED ,  RAUENBUSCH ERICH ,  ARENS ALFRED ,  IRION ECKART

IPC: C12N9/82 ,  C12N9/96 ,  C07G7/02

CPC classification number: C12N9/96 ,  C12N9/82 ,  Y10S435/816 ,  Y10S435/849

Abstract: L-asparaginase is obtained in a highly purified, pyrogen free and stabilized form from a crude aqueous solution thereof, by adding to said solution an amino acid, such as glycine, adjusting the pH of the solution to from 7.0 to 9.2, preferably in the presence of a lower aliphatic alcohol, such as methanol, then heating the solution for a period of from 1 to 5 days at a temperature up to 65* C., preferably between 40* C. to 61* C., thereby denaturizing the inactive accompanying proteins without affecting the L-asparaginase. The precipitated, denatured proteins are then removed, as by centrifuge, the enzyme enriched solution then adjusted to a pH between 4.5 and 5.5 and fractionally precipitated with a 50 percent polyethylene glycol solution, the precipitate then washed with acetone and dried.

Abstract translation: 通过向所述溶液中加入氨基酸，例如甘氨酸，将溶液的pH调节至7.0至9.2，优选地在所述溶液中，优选地在所述溶液中加入L-天冬酰胺酶， 存在低级脂族醇如甲醇，然后在高达65℃，优选40℃至61℃的温度下将溶液加热1至5天，从而使惰性 伴随蛋白质而不影响L-天冬酰胺酶。 然后通过离心除去沉淀的变性蛋白质，然后通过离心将富酶溶液调节至4.5至5.5之间的pH并用50％聚乙二醇溶液分级沉淀，然后用丙酮洗涤并干燥。

公开(公告)号：US3622461A

公开(公告)日：1971-11-23

申请号：US3622461D

申请日：1968-12-26

Applicant: BAYER AG

Inventor： WAGNER OTTO ,  BAUER KLAUS ,  KAUFMANN WILFRIED ,  RAUENBUSCH ERICH ,  ARENS ALFRED ,  IRION ECKART

IPC: C12N9/82 ,  C07G7/028

CPC classification number: C12N9/82 ,  Y10S435/816 ,  Y10S435/849

Abstract: From a culture of E. coli the bacterial mass is flocculated by addition of an organic base or a salt thereof having an amino function and a molecular weight above 1,000. The flocculated cells are centrifuged, resuspended in water and precipitated by a water-miscible solvent, such as acetone. The cells which are simultaneously opened by the previous step are centrifuged away and again resuspended in water, whereby the enzyme is extracted from the cells. By addition of the organic base, mentioned above, to the suspension, the extracted cells, nucleic acids and ballast proteins are precipitated and are removed by centrifugation. The L-asparaginase is precipitated from the cell-free solution by addition of acetone.

公开(公告)号：US3620926A

公开(公告)日：1971-11-16

申请号：US3620926D

申请日：1969-12-23

Applicant: BAYER AG

Inventor： WAGNER OTTO ,  BAUER KLAUS ,  KAUFMANN WILFRIED ,  RAUENBUSCH ERICH ,  ARENS ALFRED ,  IRION ECKART

IPC: C12N9/82 ,  C07G7/02

CPC classification number: C12N9/82 ,  Y10S435/816 ,  Y10S435/849

Abstract: L-asparaginase is obtained in concentrated purified form by fractional precipitation of an aqueous solution thereof with low molecular weight solvents, specifically diols of 4 to 8 carbon atoms, such as 2-methyl-pentane-diol-(2,4) and including such diols wherein one hydroxyl group is alkylated, for example 3methoxybutanol.