重组大环类硫肽中包含非天然氨基酸。
Recombinant Macrocyclic Lanthipeptides Incorporating Non-Canonical Amino Acids.
机构信息
The Scripps Research Institute , 10550 N Torrey Pines Road, La Jolla, California 92037, United States.
出版信息
J Am Chem Soc. 2017 Aug 30;139(34):11646-11649. doi: 10.1021/jacs.7b04159. Epub 2017 Aug 15.
Abstract
Nisin is a complex lanthipeptide that has broad spectrum antibacterial activity. In efforts to broaden the structural diversity of this ribosomally synthesized lantibiotic, we now report the recombinant expression of Nisin variants that incorporate noncanonical amino acids (ncAAs) at discrete positions. This is achieved by expressing the nisA structural gene, cyclase (nisC) and dehydratase (nisB), together with an orthogonal nonsense suppressor tRNA/aminoacyl-tRNA synthetase pair in Escherichia coli. A number of ncAAs with novel chemical reactivity were genetically incorporated into NisA, including an α-chloroacetamide-containing ncAA that allowed for the expression of Nisin variants with novel macrocyclic topologies. This methodology should allow for the exploration of lanthipeptide variants with new or enhanced activities.
摘要
尼生素是一种复杂的兰尼丁肽，具有广谱抗菌活性。为了拓宽这种核糖体合成的类细菌素的结构多样性，我们现在报告了在离散位置掺入非典型氨基酸（ncAAs）的尼生素变体的重组表达。这是通过在大肠杆菌中表达 nisA 结构基因、环化酶（nisC）和脱水酶（nisB）以及正交无意义抑制 tRNA/氨酰-tRNA 合成酶对来实现的。许多具有新颖化学反应性的 ncAAs 被遗传整合到 NisA 中，包括含有α-氯乙酰胺的 ncAA，这允许表达具有新颖的大环拓扑结构的尼生素变体。这种方法应该可以探索具有新的或增强的活性的兰尼丁肽变体。
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