大肠杆菌
黄曲霉
尿酸氧化酶
发酵
生物化学
化学
酶
药剂学
基因
生物
食品科学
药理学
作者
Jianmin Li,Zhao Chen,Lihua Hou,Huahao Fan,Shao-Jie Weng,Chun’e Xu,Jun Ren,Bing Li,Wei Chen
标识
DOI:10.1016/j.pep.2006.02.003
摘要
The entire encoding region for Aspergillus flavus uricase was cloned into pET-32a and expressed in Escherichia coli BL21 (DE3). The uricase was expressed in the E. coli cytoplasm in a completely soluble, biologically active form. A scalable process aimed to produce and purify multi-gram quantities of highly pure, recombinant urate oxidase (rUox) from E. coli was developed. The rUox protein was produced in a 30 L fermentor containing 25 L of 2× YT medium and purified to >99% purity using hydrophobic interaction, anion-exchange, and gel filtered chromatography. The final yield of purified rUox from fermentation resulted in approximately 27 g of highly pure, biologically active rUox per kg of cell paste (approximately 238 mg/8.8 g cell paste/L). The results presented here exhibit the ability to generate multi-gram quantities of rUox from E. coli that may be used for the development of pharmaceutics of reducing the hyperuricemia.
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