| 王淑婷,乔晓妮,孙晓雯,等.一株海洋源副地衣芽孢杆菌拮抗MRSA活性多肽的中试发酵工艺研究[J].中国海洋药物,2026,(3):-. |
| 一株海洋源副地衣芽孢杆菌拮抗MRSA活性多肽的中试发酵工艺研究 |
| Exploration of Pilot-Scale Fermentation Techniques for the Production of Anti-MRSA Peptide by Marine Bacillus paralicheniformis C28-2 |
| 投稿时间：2024-12-08 修订日期：2025-03-13 |
| DOI： |
| 中文关键词: 深海副地衣芽孢杆菌 抗菌肽 抗MRSA活性 发酵条件优化 中试规模工程化工艺。 |
| English Keywords:Bacillus paralicheniformis from the deep sea antibacterial peptide Activity against MRSA Optimization of fermentation Pilot scale engineering techniques. |
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| 中文摘要: |
| 目的 优化海洋源副地衣芽孢杆菌C28-2拮抗MRSA的代谢产物的发酵条件、建立工程化中试发酵工艺,以便获取足量提取物,分析鉴定抗菌活性物质的化学本质并分析该菌株的工程化潜力。方法先后采用单因素实验、Plackett-Burman、响应面优化法,研究副地衣芽孢杆菌C28-2抗菌活性物质的最适发酵条件,并完成100 L中试规模工艺的工程化放大。结果 最适发酵培养基为：大豆粉＝11.1 g/L,酪氨酸＝15.6 g/L,海水素5.0 g/L,磷酸氢二钾0.5 g/L,pH=7；摇瓶最适发酵条件：装液量,20%；接种量 ,1%；培养温度,37 ℃；转速,180 r/min；发酵周期,20 h。上述条件应用液体深层发酵,并在100 L规模建立C28-2的中试发酵工艺,成功使抑菌效价达到12800 Au/mL,比摇瓶效价提高了8倍。发酵所得的抑菌活性提取物经Tricine SDS-PAGE分析证实其为分子量在3.3 KDa右的活性多肽,为初生代谢产物。结论 海洋源副地衣芽孢杆菌C28-2能在100 L 中试规模发酵生产抗MRSA活性多肽,这个结果也揭示了该菌株表现出作为不错的工程化菌株在更大规模发酵生产抗MRSA活性多肽的潜力。 |
| English Summary: |
| Objective: Through establishing a set of Pilot scale fermentation techniques for the production of anti-MRSA peptide by the Bacillus paralicheniformis C28-2, to obtain adequate fermentate extract and check the chemical essence of antibacterial active substances, as well as to evaluate the engineering exploration potential of the strain. Method: Single factor trials, Plackett-Burman and Response surface optimization method were conducted sequentially to optimize the fermentation conditions of Bacillus paralicheniformis C28-2 at flak level, and the conditions were applied to the establishment of a set of 100 L pilot scale fermentation techniques for the production of the anti-MRSA materials. Result: The optimal fermentation medium obtained contained: soybean powder 11.1 g/L, tyrosine 15.6 g/L, seawater element 5.0 g/L, and potassium dihydrogen phosphate 0.5 g/L, pH at 7.0; The optimal fermentation conditions was as follows: volume of medium, 20%; inoculation rate, 1%; incubation temperature, 37 ℃; agitation rate, 180 r/min and cultivation time, 20 hours. By applying the conditions aforementioned, a 100 L pilot scale submerged fermentation techniques by C28-2 was successfully established which delivered a 8 folds yield improvement with antibacterial titer amounting to 12800 Au/mL. The antibacterial active substance extract derived from the fermentate was subjected to Tricine SDS-PAGE analysis, through which an anti-MRSA peptide around 3.3 KDa was identified. Conclusion: Marine B. paralicheniformis C28-2 could produce an anti-MRSA peptides at decent level via submerged fermentation process carried out at 100 L pilot scale. And this finding simply revealed the potential of C28-2 to be used as a decent engineering strain for the submerged production of anti-MRSA peptide at larger scale. |
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