Preparation method of recombinant can diad urate oxidase

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Patent number: CN1536078
Publication date: 2004-10-13
Inventor: WU YANZHUO (CN); XU MINGBO (CN); CHEN YAO (CN)
Applicant: BEIJING SHUANGLU PHARMACEUTICA (CN)
Classification:
- international: C12N9/02; C12N15/31; C12N15/53; C12N15/63; C12N15/70; C07K1/18; A61K38/44
- european:
Application number: CN20030109150 20030407
Priority number(s): CN20030109150 20030407
Abstract of CN1536078
The present invention relates to a preparation method of candida urato oxidase (UOX). It utilizes prokaryotic expression principle and technique, optimizes related expression elements of SD sequence, etc. and utilizes PCR method to clone candida (AS 2.1495) UOX gene and selectively uses pBV220 vector to construct recombinant plasmid, transforms colibacillus engineering bacterium DH5 alpha or JM109 and makes induction expression at 42 deg.C, and its expression quantity is reached to 50-60% of somatic total protein. After the engineering bacterium is undergone the processes of fermentation and bacterium breakage, the supernatant fluid can be remained, said supernatant fluid is undergone the processes of acid precipitation, cationic exchange and anion exchange treatment to obtain purified product. After the protection agent is added, it can be made into medicinal freeze-dried preparation for curing hyperuricemia disease.
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