Expression and purification of antimicrobial peptide buforin IIb in Escherichia coli
Expression and purification of antimicrobial peptide buforin IIb in Escherichia coli
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DOI：
10.1007/s10529-011-0687-4
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发表时间：
2011-07
影响因子：
2.7
通讯作者：
Qi Wang;Fenfen Zhu;Yinqiang Xin;J. Liu;L. Luo;Z. Yin
中科院分区：
文献类型：
--
作者：
Qi Wang;Fenfen Zhu;Yinqiang Xin;J. Liu;L. Luo;Z. Yin
开发了一种新的在大肠杆菌中生产21个氨基酸的抗菌肽--蟾酥蛋白IIb的方法，该蛋白是蟾酥蛋白II的合成类似物。将该基因克隆到载体pET32a中，构建了表达载体pET32a-Buforin IIb。用硝基三乙酸镍(Ni-NTA)树脂层析纯化融合蛋白Trx-Buforin IIb，经盐酸羟胺裂解后释放重组蟾毒蛋白IIb。用高效液相色谱法对重组蟾毒蛋白IIb进行了纯化，得到了纯度为99%、活性约为3.1 mg/L的重组蟾毒蛋白IIb。重组蟾毒蛋白IIb具有与人工合成的相似的抗菌活性。
A novel production method inEscherichia colifor an antimicrobial peptide of 21 amino acids, buforin IIb, which is a synthetic analog of buforin II, has been developed. The buforin IIb gene was cloned into the vector pET32a to construct an expression vector pET32a–buforin IIb. The fusion protein Trx-buforin IIb, purified by nickel nitrilo-triacetic acid (Ni-NTA) resin chromatography, was cleaved by hydroxylamine hydrochloride to release recombinant buforin IIb. Purification of recombinant buforin IIb was achieved by HPLC: about 3.1 mg/l active recombinant buforin IIb with purity >99% was obtained. The recombinant buforin IIb showed antimicrobial activities that were similar to the synthetic one.