Haoqian Liang, Youran Luo, Wilfred A. van der Donk
{"title":"Substrate Specificity of a Methyltransferase Involved in the Biosynthesis of the Lantibiotic Cacaoidin","authors":"Haoqian Liang, Youran Luo, Wilfred A. van der Donk","doi":"10.1021/acs.biochem.4c00150","DOIUrl":null,"url":null,"abstract":"Modification of the N- and C-termini of peptides enhances their stability against degradation by exopeptidases. The biosynthetic pathways of many peptidic natural products feature enzymatic modification of their termini, and these enzymes may represent a valuable pool of biocatalysts. The lantibiotic cacaoidin carries an <i>N</i>,<i>N</i>-dimethylated N-terminal amine group. Its biosynthetic gene cluster encodes the putative methyltransferase Cao4. In this work, we present reconstitution of the activity of the enzyme, which we termed CaoS<sub>C</sub> following standardized lanthipeptide nomenclature, using a heterologously produced peptide as the model substrate. In vitro methylation of diverse lanthipeptides revealed the substrate requirements of CaoS<sub>C</sub>. The enzyme accepts peptides of varying lengths and C-terminal sequences but requires dehydroalanine or dehydrobutyrine at the second position. CaoS<sub>C</sub>-mediated dimethylation of natural lantibiotics resulted in modestly enhanced antimicrobial activity of the lantibiotic haloduracin compared to that of the native compound. Improved activity and/or metabolic stability as a result of methylation illustrates the potential future application of CaoS<sub>C</sub> in the bioengineering of therapeutic peptides.","PeriodicalId":501642,"journal":{"name":"Biochemistry","volume":"51 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochemistry","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1021/acs.biochem.4c00150","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Modification of the N- and C-termini of peptides enhances their stability against degradation by exopeptidases. The biosynthetic pathways of many peptidic natural products feature enzymatic modification of their termini, and these enzymes may represent a valuable pool of biocatalysts. The lantibiotic cacaoidin carries an N,N-dimethylated N-terminal amine group. Its biosynthetic gene cluster encodes the putative methyltransferase Cao4. In this work, we present reconstitution of the activity of the enzyme, which we termed CaoSC following standardized lanthipeptide nomenclature, using a heterologously produced peptide as the model substrate. In vitro methylation of diverse lanthipeptides revealed the substrate requirements of CaoSC. The enzyme accepts peptides of varying lengths and C-terminal sequences but requires dehydroalanine or dehydrobutyrine at the second position. CaoSC-mediated dimethylation of natural lantibiotics resulted in modestly enhanced antimicrobial activity of the lantibiotic haloduracin compared to that of the native compound. Improved activity and/or metabolic stability as a result of methylation illustrates the potential future application of CaoSC in the bioengineering of therapeutic peptides.
对肽的 N 端和 C 端进行修饰可增强其稳定性,防止被外肽酶降解。许多多肽天然产物的生物合成途径都以酶修饰其末端为特征,这些酶可能是宝贵的生物催化剂库。兰特生物可可碱带有一个 N,N-二甲基化的 N-末端胺基。其生物合成基因簇编码推测的甲基转移酶 Cao4。在这项工作中,我们利用异源生产的肽作为模型底物,重组了该酶的活性,并根据标准化的anthipeptide命名法将其称为CaoSC。体外甲基化不同的anthipeptides揭示了CaoSC对底物的要求。该酶可接受不同长度和 C 端序列的肽,但在第二个位置需要脱氢丙氨酸或脱氢丁氨酸。CaoSC 介导的天然兰替霉素二甲基化使兰替霉素的抗菌活性略高于原生化合物。甲基化后活性和/或代谢稳定性的提高说明了 CaoSC 在治疗肽生物工程中的潜在应用前景。