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2025年8月3日 星期日
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重组水稻几丁质酶的发酵生产、纯化及酶学性质的研究
阎瑞香 吴仲 候建华 丁东风 李明刚
南开大学分子所 南开大学分子所
Study on the Production, Purification and Characteristics of the recombinant Chitinase from Rice
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摘要 用7L发酵罐对酵母工程菌株NB01进行了重组几丁质酶的诱导表达,通过提高通气量和调整搅拌转速,使溶氧维持在30%以上,NB01可在发酵84 h内达到产酶高峰,酶活最高可达48.0795 U/mL。利用中空纤维超滤膜对发酵液浓缩、提纯,再经过Sephadex G-100柱层析后得到单一组分的重组几丁质酶,酶纯度提高了 7.9961倍。研究结果还表明,该酶具有良好的热稳定性和耐酸性;在4℃条件下具有较好的贮藏稳定性。
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阎瑞香吴仲候建华丁东风李明刚
关键词 重组几丁质酶基因工程菌发酵纯化    
Abstract:In a 7 L fermentor, the production conditions of engineering strain NB01 containing chitinase gene were investigated. By enhancing aerati0 and changing the agitation rate,the DO level was higher than 30%. The methanol feed rate was varied with the change of DO level in order to avoid the methanol concentration too higher to injure the cells. And the results showed that the maximum chitinase activity could reach 48.0795 U/mL at 84h. The fast purification of recombinant chitinase was conducted with Hollow fiber UF membrane and Sephadex G-100 and the SDS-PAGE analysis indicated that there is a single band with the right molecular weight corresponding to the chitinase on the gel. Furthermore, the recombinant enzyme is stable at a wide range of temperature and pH. Also, it is rather stable when stored at 4℃ for a long time.
Key wordsrecombinant chitinase    engineering strain    fermentation    purification
收稿日期: 2007-06-15     
通讯作者: 李明刚   
引用本文:   
阎瑞香 吴仲 候建华 丁东风 李明刚. 重组水稻几丁质酶的发酵生产、纯化及酶学性质的研究[J]. , 2008, 16(1): 0-.
. Study on the Production, Purification and Characteristics of the recombinant Chitinase from Rice. , 2008, 16(1): 0-.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/      或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2008/V16/I1/0
 
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