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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (2): 78-83    DOI: 10.13523/j.cb.20150212
    
Pilot-scale Fermentation and Purification of a Recombinant E. coli Heat-labile Enterotoxin B Subunit
LIU Ya-long, YAN Dong-ming, WENG Liang, ZOU Xue, LIU Dan, PENG Chao, SU Ya-nan, YAN Jin-jin, ZHANG Jing, GUO Zhi-yan
Wuhu Kangwei Biontechnology Co., Ltd., Wuhu 241000, China
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Abstract  

The recombinant bacteria were induced to express Escherichia coli heat labile enterotoxin B subunit (LTB) in the form of inclsion body after pilot-scale fermentation. Q Sepharose High Perfomance Anion exchange column chromatography were used to purify the soluble protein after processing of washing and lysing, resulting to obtain the recombinant protein LTB with a high purify of 98%. Renaturation the protain through Sephadex G-25 molecular sieve chromatography to remove the salts and urea, and ultimately obtain the target protein of 95% putify. The immune double diffusion test was performed to indicate that the target protein has a good biological activity. This was the first time to determine the optimal pilot-scale fermentation conditions, the production process of E. coli LTB recombinant proteins, and also to establish the method in the use of Q column to obtain the target protein in only one step. The process was more convenient and efficient for industrial production which laid a solid foundation for the production and application of recombinant LTB protein.

Key words： LTB      Inclusion bodies      Purification     
Received: 28 October 2014      Published: 25 February 2015
ZTFLH:  Q789  
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Cite this article:

LIU Ya-long, YAN Dong-ming, WENG Liang, ZOU Xue, LIU Dan, PENG Chao, SU Ya-nan, YAN Jin-jin, ZHANG Jing, GUO Zhi-yan. Pilot-scale Fermentation and Purification of a Recombinant E. coli Heat-labile Enterotoxin B Subunit. China Biotechnology, 2015, 35(2): 78-83.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150212     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I2/78


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