Institute of Microbiology, Eidgenössische Technische Hochschule (ETH) Zürich, Vladimir-Prelog-Weg 4, 8093, Zürich, Switzerland.
Department of Pharmacy, National University of Singapore, 18 Science Drive 4, Singapore, 117543, Singapore.
Angew Chem Int Ed Engl. 2020 Nov 23;59(48):21442-21447. doi: 10.1002/anie.202008990. Epub 2020 Sep 17.
Ornithine is a component of many bioactive nonribosomal peptides but is challenging to incorporate into ribosomal products. We recently identified OspR, a cyanobacterial arginase-like enzyme that installs ornithines in the antiviral ribosomally synthesised and posttranslationally modified peptide (RiPP) landornamide A. Here we report that OspR belongs to a larger family of peptide arginases from diverse organisms and RiPP types. In E. coli, seven selected enzymes converted arginine into ornithine with little preference for the leader type. A broad range of peptide sequences was modified, including polyarginine repeats. We also generated analogues of ornithine-containing nonribosomal peptides using RiPP technology. Five pseudo-nonribosomal products with ornithines at the correct positions were obtained, including a brevicidine analogue containing ornithine and a d-amino acid installed by the peptide epimerase OspD. These results suggest new opportunities for peptide bioengineering.
鸟氨酸是许多生物活性非核糖体肽的组成部分，但很难掺入核糖体产物中。我们最近鉴定出一种蓝藻精氨酰酶样酶 OspR，它在抗病毒核糖体合成和翻译后修饰肽 (RiPP) landornamide A 中安装鸟氨酸。在这里，我们报告 OspR 属于来自不同生物体和 RiPP 类型的更大的肽精氨酸酶家族。在大肠杆菌中，七种选定的酶将精氨酸转化为鸟氨酸，对先导类型几乎没有偏好。广泛的肽序列被修饰，包括多精氨酸重复序列。我们还使用 RiPP 技术生成了含鸟氨酸的非核糖体肽类似物。获得了五个含有正确位置鸟氨酸的假非核糖体产物，包括含有鸟氨酸和由肽差向异构酶 OspD 安装的 D-氨基酸的 brevicidine 类似物。这些结果为肽的生物工程提供了新的机会。
