期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 23, 期 23, 页码 -出版社
MDPI
DOI: 10.3390/ijms232315202
关键词
antimicrobial peptide; Escherichia coli; intein; intein-mediated protein ligation
资金
- Intelligent Synthetic Biology Center of the Global Frontier Project
Antimicrobial peptides (AMPs) have been studied as potential antimicrobial coating agents, but an efficient and economical method for AMP production is lacking. Researchers successfully synthesized an AMP coating material and demonstrated its equivalent performance to peptides obtained through solid-phase synthesis.
Microbial infections remain a global health concern, calling for the urgent need to implement effective prevention measures. Antimicrobial peptides (AMPs) have been extensively studied as potential antimicrobial coating agents. However, an efficient and economical method for AMP production is lacking. Here, we synthesized the direct coating adhesive AMP, NKC-DOPA(5), composed of NKC, a potent AMP, and repeats of the adhesive amino acid 3,4-dihydroxyphenylalanine (DOPA) via an intein-mediated protein ligation strategy. NKC was expressed as a soluble fusion protein His-NKC-GyrA (HNG) in Escherichia coli, comprising an N-terminal 6x His-tag and a C-terminal Mxe GyrA intein. The HNG protein was efficiently produced in a 500-L fermenter, with a titer of 1.63 g/L. The NKC-thioester was released from the purified HNG fusion protein by thiol attack and subsequently ligated with chemically synthesized Cys-DOPA(5). The ligated peptide His-NKC-Cys-DOPA(5) was obtained at a yield of 88.7%. The purified His-NKC-Cys-DOPA(5) possessed surface-binding and antimicrobial properties identical to those of the peptide obtained via solid-phase peptide synthesis. His-NKC-Cys-DOPA(5) can be applied as a practical and functional antimicrobial coating to various materials, such as medical devices and home appliances.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据