Exploring bioactive peptides from bacterial secretomes using PepSAVI-MS: identification and characterization of Bac-21 from Enterococcus faecalis pPD1.

Exploring bioactive peptides from bacterial secretomes using PepSAVI-MS: identification and characterization of Bac-21 from Enterococcus faecalis pPD1.
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DOI:
10.1111/1751-7915.13299
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发表时间:
2018-09
影响因子:
5.7
通讯作者:
Hicks LM
Hicks LM
中科院分区:
工程技术2区
文献类型:
--
作者:
Kirkpatrick CL;Parsley NC;Bartges TE;Wing CE;Kommineni S;Kristich CJ;Salzman NH;Patrie SM;Hicks LM

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由于抗生素耐药性的上升正在超过目前的抗生素药物发现方法,因此需要新方法和创新技术来补充我们不断减少的抗菌剂库。为此,我们开发了PepSAVI‐MS管道,以加快对天然产物生物活性肽的搜索。在本文中,我们通过鉴定来自粪肠球菌pPD 1的细菌素Bac-21,证明了PepSAVI-MS用于发现细菌来源的生物活性肽的扩展。微小的管道修改,包括实施细菌注入琼脂扩散测定和可选的肽库消化,突出了PepSAVI-MS管道的多功能性和广泛的适应性。此外,我们首次通过实验验证了活性成熟Bac-21肽的一级蛋白质序列,并证实了其在一级序列和翻译后加工方面的同一性。如本文所证明的,PepSAVI-MS成功应用于细菌分泌物组建立了用于新型微生物生物活性肽发现的原理证明。
As current methods for antibiotic drug discovery are being outpaced by the rise of antimicrobial resistance, new methods and innovative technologies are necessary to replenish our dwindling arsenal of antimicrobial agents. To this end, we developed the PepSAVI‐MS pipeline to expedite the search for natural product bioactive peptides. Herein we demonstrate expansion of PepSAVI‐MS for the discovery of bacterial‐sourced bioactive peptides through identification of the bacteriocin Bac‐21 from Enterococcus faecalis pPD1. Minor pipeline modifications including implementation of bacteria‐infused agar diffusion assays and optional digestion of peptide libraries highlight the versatility and wide adaptability of the PepSAVI‐MS pipeline. Additionally, we have experimentally validated the primary protein sequence of the active, mature Bac‐21 peptide for the first time and have confirmed its identity with respect to primary sequence and post‐translational processing. Successful application of PepSAVI‐MS to bacterial secretomes as demonstrated herein establishes proof‐of‐principle for use in novel microbial bioactive peptide discovery.
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通讯作者: Hicks LM