Sethi Harkirat S, Osier Jessica L, Burks Geordan L, Lamar Jennifer F, McFeeters Hana, McFeeters Robert L
Department of Chemistry, University of Alabama in Huntsville, Huntsville, AL, USA.
AIMS Mol Sci. 2017;4(2):175-184. doi: 10.3934/molsci.2017.2.175. Epub 2017 May 12.
New antibiotics and new antibiotic targets are needed to counter the development of bacterial drug resistance that threatens to return the human population to the pre-antibiotic era. Bacterial peptidyl-tRNA hydrolase (Pth1) is a promising new antibiotic target in the early stages of development. While inhibitory activity has been observed in a variety of natural products, bioactive fractionation has been a bottleneck for inhibitor isolation. To expedite the isolation of inhibitory compounds from complex mixtures, we constructed a Pth1 affinity column and used it to isolate inhibitory compounds from crude natural products. Recombinantly produced Pth1 was covalently attached to a column matrix and the inhibitory activity isolated from ethanol extracts of . The procedure reported here demonstrates that isolation of Pth1 inhibitory compounds from complex natural product extracts can be greatly expedited over traditional bioactive fractionation, decreasing time and expense. The approach is generally applicable to Pth1s from other bacterial species and opens an avenue to advance and accelerate inhibitor development against this promising antimicrobial target.
为应对细菌耐药性的发展，需要研发新的抗生素和新的抗生素靶点，否则人类可能会重回抗生素出现之前的时代。细菌肽基 - tRNA水解酶（Pth1）是处于研发早期阶段的一个很有前景的新抗生素靶点。虽然在多种天然产物中已观察到抑制活性，但生物活性分级分离一直是抑制剂分离的瓶颈。为了加快从复杂混合物中分离抑制性化合物的速度，我们构建了一个Pth1亲和柱，并使用它从天然产物粗提物中分离抑制性化合物。重组产生的Pth1被共价连接到柱基质上，从[具体来源]的乙醇提取物中分离出抑制活性。本文报道的方法表明，与传统的生物活性分级分离相比，从复杂天然产物提取物中分离Pth1抑制性化合物的速度可大大加快，从而减少时间和费用。该方法一般适用于其他细菌物种的Pth1，并为推进和加速针对这一有前景的抗菌靶点的抑制剂开发开辟了一条途径。
