期刊
JOURNAL OF BIOTECHNOLOGY
卷 234, 期 -, 页码 83-89出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2016.07.021
关键词
ELP; Intein; Antimicrobial peptide; Batch production
资金
- National Council of Technological and Scientific Development (CNPq)
- Coordination for the Improvement of Educational Personnel (CAPES)
- Foundation for Research Support of the Federal District (FAPDF)
- Foundation for Educational Research and Technology of Mato Grosso do Sul (FUNDECT)
- Ministry of Science and Technology (MCT)
An important aspect related to infectious pathogens is their exceptional adaptability in developing resistance, which leads to a perpetual challenge in the discovery of antimicrobial drugs with novel mechanisms of action. Among them, antimicrobial peptides (AMPs) stand out as promising anti-infective molecules. In order to overcome the high costs associated with isolation from natural sources or chemical synthesis of AMPs we propose the expression of Pa-MAP 2, a polyalanine AMP. Pa-MAP 2 was fused to an ELP-intein tag where the ELP (Elastin-like polypeptide) was used to promote aggregation and fast and cost-effective isolation after expression, and the intein was used to stimulate a controlled AMP release. For these, the vector pET2la was used to produce Pa-MAP 2 fused to the N-termini region of a modified Mxe GyrA intein followed by 60 repetitions of ELP. Purified Pa-MAP 2 showed a MIC of 25 tilVI against E. coli ATCC 8739. Batch fermentation demonstrated that Pa-MAP-2 can be produced in both rich and defined media at yields 50-fold higher than reported for other AMPs produced by the ELP-intein system, and in comparable yields to expression systems with protease or chemical cleavage. (C) 2016 Elsevier B.V. All rights reserved.
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