Stachelhaus T, Schneider A, Marahiel M A
Biochemie/Fachbereich Chemie, Philipps-University of Marburg, Germany.
Science. 1995 Jul 7;269(5220):69-72. doi: 10.1126/science.7604280.
Peptide synthetases involved in the nonribosomal synthesis of peptide secondary metabolites possess a highly conserved domain structure. The arrangement of these domains within the multifunctional enzymes determines the number and order of the amino acid constituents of the peptide product. A general approach has been developed for targeted substitution of amino acid-activating domains within the srfA operon, which encodes the protein templates for the synthesis of the lipopeptide antibiotic surfactin in Bacillus subtilis. Exchange of domain-coding regions of bacterial and fungal origin led to the construction of hybrid genes that encoded peptide synthetases with altered amino acid specificities and the production of peptides with modified amino acid sequences.
参与肽类次生代谢产物非核糖体合成的肽合成酶具有高度保守的结构域结构。这些结构域在多功能酶中的排列决定了肽产物中氨基酸组成成分的数量和顺序。已开发出一种通用方法，用于对枯草芽孢杆菌中编码脂肽抗生素表面活性素合成的蛋白质模板的srfA操纵子内的氨基酸激活结构域进行靶向替换。细菌和真菌来源的结构域编码区域的交换导致构建了编码具有改变的氨基酸特异性的肽合成酶的杂合基因，并产生了具有修饰氨基酸序列的肽。
