Zorko Mateja, Jerala Roman
Department of Biotechnology, National Institute of Chemistry, Ljubljana, Slovenia.
Methods Mol Biol. 2010;618:61-76. doi: 10.1007/978-1-60761-594-1_5.
Large quantities of antimicrobial peptides are required for investigations and clinical trials, therefore suitable production method alternative to traditional chemical synthesis is necessary. Production of recombinant antimicrobial peptides in prokaryotic systems has successfully demonstrated the viability of this approach. Production of antimicrobial peptides in Escherichia coli is potentially limited due to their toxicity to host cells and susceptibility to proteolytic degradation, which can be avoided using fusion protein approach. We describe antimicrobial peptide production in E. coli based on forcing antimicrobial peptides into inclusion bodies, which is affective for the production of large quantities of antimicrobial peptides. Chemical reagents for cleaving peptide bond between antimicrobial peptides and fusion proteins such as cyanogen bromide and diluted acid are selective and provide antimicrobial peptides for biological studies in short time.
研究和临床试验需要大量的抗菌肽，因此有必要采用替代传统化学合成的合适生产方法。在原核系统中生产重组抗菌肽已成功证明了这种方法的可行性。由于抗菌肽对宿主细胞有毒性且易受蛋白水解降解，在大肠杆菌中生产抗菌肽可能会受到限制，而使用融合蛋白方法可以避免这种情况。我们描述了基于将抗菌肽强制纳入包涵体的大肠杆菌中抗菌肽的生产，这对于大量生产抗菌肽是有效的。用于切割抗菌肽与融合蛋白之间肽键的化学试剂，如溴化氰和稀酸，具有选择性，并能在短时间内为生物学研究提供抗菌肽。
