Müller Hagen, Salzig Denise, Czermak Peter
Inst. of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, Giessen, 35390, Germany.
Biotechnol Prog. 2015 Jan-Feb;31(1):1-11. doi: 10.1002/btpr.2002. Epub 2014 Oct 28.
Antimicrobial peptides (AMPs) could evolve into new therapeutic lead molecules against multi-resistant bacteria. As insects are a rich source of AMP, the identification and characterization of insect-derived AMPs is particularly emphasized. One challenge of bringing these molecules into market, e.g., as a drug, is to develop a cost-efficient large-scale production process. Due to the fact that a direct AMP isolation from insects is not economical and that chemical synthesis is recommended for peptide sizes below 40 amino acids, a viable option is heterologous AMP production. Therefore, previous knowledge concerning the expression of larger proteins can be adapted, but due to the AMP nature (e.g., small size, bactericide) additional challenges have to be faced during up and downstream processing. Nonetheless the bottleneck for large-scale AMP production is the same as for proteins; mainly the downstream process. This review introduces opportunities for insect-derived AMP production, like the choice of the expression system (based on previously derived data), depending on the AMP nature, as well as new purification strategies like elastin-like peptide/intein based purification strategies. All of these aspects are discussed with regard to large-scale processes and costs.
抗菌肽(AMPs)有望发展成为对抗多重耐药菌的新型治疗先导分子。由于昆虫是抗菌肽的丰富来源,因此特别强调对昆虫源抗菌肽的鉴定和表征。将这些分子推向市场(例如作为药物)面临的一个挑战是开发一种经济高效的大规模生产工艺。由于直接从昆虫中分离抗菌肽不经济,且对于40个氨基酸以下的肽推荐采用化学合成法,因此一个可行的选择是异源生产抗菌肽。因此,可以借鉴以往关于较大蛋白质表达的知识,但由于抗菌肽的特性(例如,小分子、杀菌作用),在上游和下游加工过程中还必须面对其他挑战。尽管如此,大规模生产抗菌肽的瓶颈与生产蛋白质的瓶颈相同;主要是下游加工过程。本综述介绍了昆虫源抗菌肽生产的机遇,如根据抗菌肽的特性选择表达系统(基于先前获得的数据),以及基于弹性蛋白样肽/内含肽的纯化策略等新的纯化策略。所有这些方面都结合大规模生产工艺和成本进行了讨论。