Evolution-guided engineering of nonribosomal peptide synthetase adenylation domains
Evolution-guided engineering of nonribosomal peptide synthetase adenylation domains
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DOI：
10.1039/c2sc21722h
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发表时间：
2013-01-01
期刊：
影响因子：
8.4
通讯作者：
Piel, Joern
中科院分区：
文献类型：
--
作者：
Cruesemann, Max;Kohlhaas, Christoph;Piel, Joern
荷霉素是一种由细菌非核糖体肽合成酶（NRPS）生物合成的结构异常的形态发生和抗生素肽。生物信息学分析表明，部分NRPS腺苷酸化（A）结构域在进化过程中发生了重组，导致底物特异性的重大转变。这一特征启发我们根据假定的重组点创建具有改变的底物的A结构域。在表征所有天然的标准霉素A结构域之后，构建并表征工程化版本。三种酶显示出几乎相同的特异性概况的天然结构域识别相同的基板。这些数据支持进化假说的出现hormaomycin途径，并建议在NRPS工程的新策略。
Hormaomycin is a structurally unusual morphogenic and antibiotic peptide biosynthesized by a bacterial nonribosomal peptide synthetase (NRPS). Bioinformatic analysis suggested that parts of the NRPS adenylation (A) domains had recombined during evolution, resulting in a major switch of substrate specificity. This feature inspired us to create A domains with altered substrates based on the putative recombination points. Following characterization of all native hormaomycin A domains, engineered versions were constructed and characterized. Three of the enzymes displayed an almost identical specificity profile to that of native domains recognizing the same substrates. The data support the evolutionary hypothesis regarding the emergence of the hormaomycin pathway and suggest new strategies in NRPS engineering.