扩展和的遗传密码以纳入非天然氨基酸用于生产修饰的羊毛硫抗生素。
Expanding the Genetic Code of and to Incorporate Non-canonical Amino Acids for Production of Modified Lantibiotics.
作者信息
Bartholomae Maike, Baumann Tobias, Nickling Jessica H, Peterhoff David, Wagner Ralf, Budisa Nediljko, Kuipers Oscar P
机构信息
Department of Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, Netherlands.
Biocatalysis Group, Department of Chemistry, Technische Universität Berlin (Berlin Institute of Technology), Berlin, Germany.
出版信息
Front Microbiol. 2018 Apr 6;9:657. doi: 10.3389/fmicb.2018.00657. eCollection 2018.
The incorporation of non-canonical amino acids (ncAAs) into ribosomally synthesized and post-translationally modified peptides, e.g., nisin from the Gram-positive bacterium , bears great potential to expand the chemical space of various antimicrobials. The ncAA -Boc-L-lysine (BocK) was chosen for incorporation into nisin using the archaeal pyrrolysyl-tRNA synthetase-tRNA pair to establish orthogonal translation in for read-through of in-frame amber stop codons. In parallel, recombinant nisin production and orthogonal translation were combined in cells. Both organisms synthesized bioactive nisin(BocK) variants. Screening of a nisin amber codon library revealed suitable sites for ncAA incorporation and two variants displayed high antimicrobial activity. Orthogonal translation in and presents a promising tool to create new-to-nature nisin derivatives.
将非标准氨基酸（ncAAs）掺入核糖体合成和翻译后修饰的肽中，例如革兰氏阳性细菌中的乳链菌肽，对于扩展各种抗菌剂的化学空间具有巨大潜力。选择ncAA -Boc-L-赖氨酸（BocK），利用古菌吡咯赖氨酸-tRNA合成酶-tRNA对将其掺入乳链菌肽中，以在大肠杆菌中建立正交翻译，用于通读框内琥珀色终止密码子。同时，在大肠杆菌细胞中结合了重组乳链菌肽生产和正交翻译。两种生物体都合成了具有生物活性的乳链菌肽（BocK）变体。对乳链菌肽琥珀密码子文库的筛选揭示了适合掺入ncAA的位点，并且两个变体表现出高抗菌活性。在大肠杆菌和枯草芽孢杆菌中的正交翻译是创造新型天然乳链菌肽衍生物的有前途的工具。
Zotero 插件