生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (2): 46-.doi: 10.3969/j.issn.2095-1736.2025.02.046

• 研究报告 • 上一篇    下一篇

应用离子束注入技术改善乳杆菌维生素K2 合成能力

沐璟玮1， 杨　旭2， 汪洪波2， 王　丽3，4， 孙曼曼3， 王　鹏3   

  1. 1. 安徽大学 物质科学与信息技术研究院， 合肥 230601； 2. 安徽新希望白帝乳业有限公司，
    合肥 230600； 3. 中国科学院合肥物质科学研究院 智能机械研究所， 合肥 230031；
    4. 中国科学技术大学 研究生院科学岛分院， 合肥 230026
  • 出版日期:2025-04-18 发布日期:2025-04-14
  • 通讯作者: 王鹏，研究员，主要从事合成生物学研究，E-mail:pengwang@ipp.ac.cn
  • 作者简介:沐璟玮，硕士研究生，研究方向为合成生物学，E-mail:923183023@qq.com
  • 基金资助:
    国家自然科学基金面上项目(12375352)；安徽省科技重大专项（202103a06020003）

Application of ion beam implantation technology to improve the vitamin K2 synthesis capacity in Lactobacillus

MU Jingwei1, YANG Xu2, WANG Hongbo2, WANG Li3,4, SUN Manman3, WANG Peng3   

  1. 1. Institutes of Physical Science and Information Technology, Anhui University, Hefei 230601, China; 2. Anhui New
    Hope Baidi Dairy Co., Ltd., Hefei 230600, China; 3. Institute of Intelligent Machinery, Hefei Institutes of Physical
    Science, Chinese Academy of Sciences, Hefei 230031, China; 4. Science Island Branch of Graduate School,
    University of Science and Technology of China, Hefei 230026, China
  • Online:2025-04-18 Published:2025-04-14

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摘要： 维生素K2（VK2）对维持人体骨骼、心血管健康具有重要作用，但由于乳酸菌（LAB）VK2合成路径中部分基因沉默或不表达，导致目前乳制品中基本不含有VK2。以发酵乳杆菌Lactobacillus fermentum（LF1）为出发菌株，分析不同离子束注入条件下的致死率和正负突变率，确定最佳注入能量为10 keV、注入剂量为75（×2.6×1013 N+/cm2）。通过多轮离子束注入结合适应性实验室进化（ALE），获得VK2高产菌株LFX，产量为0.67 mg/g，并在连续传代实验中验证菌株良好的遗传稳定性。转录组分析发现，在LFX的VK2相关合成途径中，甲萘醌主环与聚异戊二烯侧链合成相关基因的表达水平显著增加，抗氧化系统、辅因子合成及中央碳氮代谢等途径显著增强。结果表明，多轮离子束注入和ALE可以协同优化代谢通量，提高VK2的生物合成和稳定性。

关键词: 离子束, 诱变选育, 适应性实验室进化, 维生素K2, 发酵乳杆菌, 转录组

Abstract: Vitamin K2 (VK2) plays a essential role in maintaining human bone and cardiovascular health. However, due to the silence or non-expression of some genes in the VK2 synthesis pathway of lactic acid bacteria, VK2 is negligible in dairy products. In this study, using Lactobacillus fermentum (LF1) as the starting strain, the lethality and the positive and negative mutation rates were analyzed under different ion beam injection conditions. The optimal parameters were determined to be an energy of 10 keV and a dose of 75(×2.6×1013 N+/cm2). Through successive ion beam iterations coupled with adaptive laboratory evolution (ALE), the high-yield VK2 strain LFX was obtained, achieving a production rate of 0.67 mg/g. The strain’s genetic stability was confirmed through serial passage experiments. Transcriptome analysis revealed that in the VK2-related synthesis pathway of LFX, genes associated with the biosynthesis of the naphthoquinone core and the polyprenyl side chain were markedly upregulated. Moreover, pathways involved in the antioxidant defense, cofactor biosynthesis, and central carbon and nitrogen metabolism were substantially enhanced. The findings suggested that iterative ion beam implantation and ALE could synergistically optimize metabolic fluxes, enhancing VK2 biosynthesis and stability in the developed strains.

Key words: ion beam, mutagenesis breeding, adaptive laboratory evolution, vitamin K2;lactobacillus fermentum, transcriptome

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