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      2. 羊毛硫抗生素雷可肽同源基因簇的異源表達
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        國家自然科學基金項目(31770036)


        Heterologous expression of lantibiotic lexapeptide biosynthetic gene cluster homologue
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          摘要:

          高杰氏鏈霉菌中含有新型羊毛硫抗生素雷可肽(lexapeptide)的同源基因簇lxm2,為探索該基因簇相應的產物,將包含lxm2的細菌人工染色體轉到變鉛青鏈霉菌中進行異源表達,證實其可以產生雷可肽;同時還獲得另一個新次級代謝產物,基因敲除實驗表明此化合物的合成也與lxm2相關。通過色譜技術對新化合物進行分離純化,結合核磁共振譜、高分辨質譜和二級質譜確定其結構為具有多重修飾的線性六肽,與雷可肽的N-端六氨基酸序列相同,命名為Lxm-N-六肽,該化合物沒有檢測到抗菌活性。

          Abstract:

          Lantibiotics attract extensive attention in the field of discovery and development of antibiotics due to its special action mode and barely observed drug-resistant mutation after being applied for decades. Lexapeptide is a type V lantibiotic recently uncovered via a functional genome mining approach (LEXAS) and has strong antibacterial activity against Gram positive bacteria including MRSA and MRSE. A gene cluster (lxm2) similar to the lexapeptide biosynthetic gene cluster was identified through screening the Streptomyces galtieri genomic DNA bacterial artificial chromosome (BAC) library via LEXAS. The BAC clone carrying lxm2 was introduced into Streptomyces lividians for heterologous expression,leading to the production of lexapeptide and a novel peak in HPLC. The production of lexapeptide and the new peak was disappeared when the lxm2 gene cluster was removed from the BAC clone. Based on NMR spectrometry and high resolution mass spectrometry,the structure of the new compound was identified to be a highly modified linear hexapeptide with sequence identical to the N-terminal six amino acids of lexapeptide,thus Lxm-N-hexapeptide was given as its name. The Lxm-N-hexapeptide did not exhibit antibacterial activity.

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        徐利軍,王業民,徐敏,趙志龍,高貴喜,陶美鳳.羊毛硫抗生素雷可肽同源基因簇的異源表達[J].華中農業大學學報,2020,39(5):93-100

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        • 收稿日期:2020-06-23
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        • 在線發布日期: 2020-10-05
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