一个基于血解素分泌途径的新型分泌表达平台,用于在大肠杆菌中高效制造标记和抗菌
Wen Zhu1, Lifu Hu1, Yang Wang1
1Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, China.
Bioresources and bioprocessing
|April 23, 2024
概括
这项研究引入了一种新的大肠杆菌分泌平台THHly,用于高效生产标签和抗菌. 该HlyA系统使直接的细胞外运输,减少毒性,并使工业制造.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 蛋白质表达方式
背景情况:
- 在大肠杆菌中的分泌表达对外源蛋白质仍然具有挑战性.
- 血清素A (HlyA) 分泌系统提供直接的细胞质到细胞外传输,绕过周等离子体中间体.
- HlyA是开发高效的秘生产平台的理想候选人.
研究的目的:
- 开发一种基于大肠杆菌HlyA系统的新型分泌物生产平台THHly.
- 探索平台在制备和检测标记和抗菌方面的效率.
- 评估分泌的抗微生物的生物活性和宿主毒性.
主要方法:
- 设计了一种融合蛋白质策略,使用HlyA信号序列,标,烟草蚀刻病毒 (TEV) 蛋白酶分裂部位和6*His标签.
- 在大肠杆菌BL21 (DE3) 中表达的融合蛋白.
- 使用亲和染色学和TEV裂变净化;评估生物活性和宿主毒性.
主要成果:
- 成功实现了五个标签的分泌表达,T7和S标签被净化并保持免疫性.
- 生产了Cecropin A和Melittin,证明了抗微生物和抗瘤生物活性.
- 融合蛋白表达没有显著抑制细菌生长,这表明抗微生物对宿主毒性降低.
结论:
- 基于HlyA的THHly平台能够有效地产生分泌物和净化.
- 这种新的平台显示了工业规模制造类药物和试剂的巨大潜力.
- 这项研究代表了第一份关于大肠杆菌中特定抗微生物的分泌表达的报告,具有工业潜力.
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