枯草芽孢杆菌SHT-15抗菌脂肽分离鉴定及对厚皮甜瓜贮藏品质的影响

Isolation and identification of antimicrobial lipopeptide from Bacillus subtilis SHT-15 and its effect on storage quality of thick-skinned melons

  • 摘要:
    目的 对枯草芽孢杆菌(Bacillus subtilis)菌株SHT-15产生的脂肽进行分离、纯化与鉴定,以及探究其对厚皮甜瓜贮藏品质的影响。
    方法 通过有机溶剂萃取法对枯草芽孢杆菌 SHT-15 脂肽进行提取,利用 SephadexG-25 葡聚糖凝胶对脂肽进行分离纯化,液相色谱-质谱联用仪(LC-MS)对纯化后的脂肽进行分析鉴定,同时在西州蜜25号厚皮甜瓜上施加该菌提取的脂肽物质,测定厚皮甜瓜贮藏期间脂氧合酶(Lipoxygenase,LOX)、乙醇脱氢酶(Alcohol dehydrogenase,ADH)的活性、糖酸比和果皮色度。
    结果 菌株SHT-15脂肽提取物对病原菌有明显抑菌作用,抑菌圈为(20.12 ± 0.54) mm,抑菌活性为 70.18 %。脂肽粗提物经过 SephadexG-25 葡聚糖凝胶层析后,洗脱曲线中共有 7 个脂肽峰,主要起抑菌作用的为第2个脂肽峰,抑菌圈为(21.44 ± 0.13) mm,抑菌活性为 72.01%。LC-MS分析表明,分离出 31 个抑菌肽段中,保留时间在 47.84 min 的肽段对应离子流图中总离子强度最高的峰,此时 m/z 值为 1046.58,当 m/z 值在 1032.4 ~1074.4 范围时属于表面活性素家族(Surfactin)。抑菌物质能提高厚皮甜瓜贮藏期LOX、ADH的活性,抑制糖酸比的下降,对维持果皮色度有较好效果。
    结论 菌株 SHT-15中主要起抑菌活性成分是 C12-C15 Surfactin,能较好地维持甜瓜贮藏期品质。

     

    Abstract:
    Objective To isolate, purify and characterize the lipopeptides produced by Bacillus subtilis strain SHT-15, and to investigate their effects on the storage quality of thick-skinned melons.
    Methods The lipopeptide of Bacillus subtilis SHT-15 was extracted by organic solvent extraction, and separated and purified by SephadexG-25 dextran gel, and analyzed and identified by liquid chromatography-mass spectrometry (LC-MS). The activities of lipoxygenase (LOX), ethanol dehydrogenase (ADH), sugar-acid ratio and rind color were determined during storage.
    Results The lipopeptide extract of strain SHT-15 had obvious inhibitory effect on the pathogenic bacteria, the inhibition circle was (20.12 ± 0.54) mm, and the inhibitory activity was 70.18%. After SephadexG-25 dextran gel chromatography, there were 7 lipopeptide peaks in the elution curve, and the main inhibitory effect was the 2nd lipopeptide peak, with a circle of inhibition of (21.44 ± 0.13) mm, and an inhibitory activity of 72.01 %. LC-MS analysis showed that, of the 31 inhibitory peptides isolated, the one with a retention time of 47.84 min corresponded to the peak with the highest total ionic strength in the ion-current diagram. The m/z value was 1046.58, and the peptide with retention time of 47.84 min belonged to the Surfactin family when the m/z value was in the range of 1032.4 to 1074.4. The bacteriostatic substances can improve the activity of LOX and ADH during the storage period of thick-skinned melons, inhibit the decline of sugar-acid ratio, and have a better effect on maintaining the color of the rind.
    Conclusion The main component of bacteriostatic activity in strain SHT-15 is C12-C15 Surfactin, which can better maintain the quality of melon during storage.

     

/

返回文章
返回