食品科学 ›› 2025, Vol. 46 ›› Issue (23): 139-149.doi: 10.7506/spkx1002-6630-20250610-062

• 生物工程 • 上一篇    

产抗菌脂肽贝莱斯芽孢杆菌的诱变育种、发酵优化及基因差异分析

方茵,宋文娇,徐铮,续晓琪,王大毛,李莎,徐虹   

  1. (1.南京工业大学食品与轻工学院,江苏 南京 211816;2.西南大学食品科学学院,重庆 400715)
  • 发布日期:2025-12-26
  • 基金资助:
    国家自然科学基金面上项目(22178171;32172158)

Mutation Breeding and Optimization of Fermentation Conditions for Lipopeptide Production by Bacillus velezensis and Differential Genomic Analysis

FANG Yin, SONG Wenjiao, XU Zheng, XU Xiaoqi, WANG Damao, LI Sha, XU Hong   

  1. (1. College of Food and Light Industry, Nanjing Tech University, Nanjing 211816, China;2. School of Food Science, Southwest University, Chongqing 400715, China)
  • Published:2025-12-26

摘要: 以前期筛选的产脂肽贝莱斯芽孢杆菌BVQ121为研究对象,通过常压室温等离子体-LiCl复合诱变技术成功选育出高产抗菌脂肽突变株BV227,结合发酵优化,脂肽产量较原始菌株BVQ121提高了122.31%。进一步通过液相色谱-质谱分析、全基因组与转录组分析发现:1)突变株可产生更多芬荠素(Fengycin)类脂肽同系物;2)4 个关键非同义单核苷酸多态性变异显著影响脂肽合成途径;3)群体感应通路基因显著富集;4)乙醇脱氢酶基因和脂肪酸羟化酶的相关合成基因显著上调,为脂肪酸链合成提供更多前体并增加了脂肽结构多样性和生物活性。随后,牛津杯实验验证诱变后菌株的脂肽对沙门氏菌和鮰鱼爱德华氏菌两种致病菌均具有明显的抑菌效果,且具有良好的热稳定性。本研究通过复合诱变和发酵优化提高了贝莱斯芽孢杆菌BVQ121的脂肽产量,验证了基因组变异对合成抗菌脂肽关键基因表达的影响,为构建基因表达调控模型、设计理性代谢工程策略提供了分子基础,同时为其在食品防腐等领域的规模化应用奠定理论依据。

关键词: 常压室温等离子体-LiCl复合诱变;贝莱斯芽孢杆菌;抗菌脂肽;基因组分析

Abstract: A mutant strain named BV227 with higher yield of antimicrobial lipopeptides was obtained from Bacillus velezensis BVQ121, a strain producing lipopeptides, by atmospheric and room temperature plasma (ARTP)-LiCl mutagenesis. Under optimized fermentation conditions, BV227 produced 122.31% more lipopeptides than did BVQ121. Liquid chromatography-mass spectrometry (LC-MS), whole-genome sequencing, and transcriptome analysis revealed that: 1) the mutant strain produced more types and larger amounts of fengycin homologs; 2) four key nonsynonymous single-nucleotide polymorphisms (SNPs) significantly affected the lipopeptide biosynthetic pathway; 3) genes involved in quorum sensing were significantly enriched; and 4) the alcohol dehydrogenase and fatty acid hydroxylase genes were significantly up-regulated, enhancing fatty acid precursor supply and increasing the structural diversity and bioactivity of lipopeptide. Antimicrobial assays using the Oxford cup method confirmed that the lipopeptides produced by the mutant strain exhibited strong inhibitory effects against Salmonella sp. and Edwardsiella ictaluri, and demonstrated excellent thermal stability. In summary, the mutant strain, in comparison to the parental strain, had an enhanced ability to produce lipopeptides under optimized fermentation conditions. Our findings confirmed the influence of genomic variation on the expression of key genes related to the synthesis of antimicrobial lipopeptides. This study provides a molecular foundation for establishing gene expression regulatory networks and rationally designing metabolic engineering strategies, and also offers a theoretical basis for the large-scale application of B. velezensis in food preservation and other fields.

Key words: atmospheric and room temperature plasma-LiCl mutagenesis; Bacillus velezensis; antimicrobial lipopeptides; genomic analysis

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