Expression, purification, and characterization of recombinant human keratinocyte growth factor-2 in Pichia pastoris
Expression, purification, and characterization of recombinant human keratinocyte growth factor-2 in Pichia pastoris
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DOI：
10.1016/j.jbiotec.2007.08.024
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发表时间：
2007-10-15
影响因子：
4.1
通讯作者：
Zhang Zhaoshan
中科院分区：
文献类型：
--
作者：
Wang Yanchun;Yuan Shengling;Zhang Zhaoshan
角质细胞生长因子-2（KGF-2）是成纤维细胞生长因子家族的成员。通过合成获得的全长人KGF-2编码序列在AOX启动子的转录控制下与酵母α-因子分泌信号框内克隆到pPICZ α A载体中，并整合到巴斯德毕赤酵母菌株GS 115中。在甲醇诱导的摇瓶培养中，rhKGF-2的含量约占总分泌蛋白的17.5%。在此条件下，rhKGF-2的产量稳定在1.0g/l左右。重组蛋白经肝素亲和层析纯化。通过Western blot分析和生物学活性测定对纯化的rhKGF-2进行了初步的生物学特性鉴定，结果表明重组KGF-2得到了成功表达。(C)2007 Elsevier B. V.保留所有权利。
Keratinocyte growth factor-2 (KGF-2) is a member of the fibroblast growth factor family. The full-length human KGF-2 coding sequence, gained by synthesizing, was cloned into the pPICZ alpha A vector in frame with the yeast alpha-factor secretion signal under the transcriptional control of the AOX promoter and integrated into Pichia pastoris strain GS115. In shake-flask culture induced with methanol, the rhKGF-2 content was about 17.5% of the total secreted proteins. Under the optimal conditions, stable production of rhKGF-2 around 1.0 g/l was achieved. The recombinant protein was purified by heparin affinity chromatography. A preliminary biochemical characterization of purified rhKGF-2 was performed both by Western blot analysis and biological activity analysis, and the result demonstrated that the recombinant KGF-2 was expressed successfully. (C) 2007 Elsevier B.V. All rights reserved.