解淀粉芽孢杆菌抑菌肽的分离鉴定及其抑菌谱表征

doi:10.7506/spkx1002-6630-20190505-020

摘要/Abstract

摘要： 以金黄色葡萄球菌为指示菌，对诱变优化总抑菌活性提高后的解淀粉芽孢杆菌发酵产物，建立酸沉、有机溶剂萃取，DEAE-Sepharose-Fast Flow阴离子交换层析，反相高效液相分离、纯化方案。对纯化产物通过液相色谱-质谱分析发现两种抑菌成分，一种为已见报道的含有C13～C15的Surfactin同系物；另一种为氨基酸序列为L-V-N-P-P-T，分子质量为639.75 Da的抑菌肽。经ProtParam软件分析，该6肽的等电点为5.52，GRAVY为0.100，脂溶性指数为113.33。将此6肽与APD数据库已报道的抑菌肽对比，同源性最高的仅为37.5%，由此推断此6肽为新型抗菌肽。对Surfactin与6肽的抑菌谱分析，发现二者对革兰氏阳性菌、革兰氏阴性菌均有抑菌作用。

关键词: 解淀粉芽孢杆菌；抑菌活性；分离；纯化；抑菌肽

Abstract: Antibacterial substances against Staphylococcus aureus from the fermentation supernatant of Bacillus amyloliquefaciens with improved antibacterial activity were separated and purified by sequential acid precipitation, organic solvent extraction, DEAE anion exchange chromatography and reversed phase-high performance liquid chromatography (RP-HPLC). Two antibacterial peptides were found in the purified product by liquid chromatography and mass spectrometry (LC-MS). One was a reported Surfactin homologue containing 13–15 carbon atoms; the other was a bacteriostatic peptide having the amino acid sequence of L-V-N-P-P-T with a molecular mass of 639.75 Da. The hexapeptide had an isoelectric point of 5.52, a GRAVY of 0.100, and a fat solubility index of 113.33 as analyzed by ProtParam software. The highest homology between this peptide with the bacteriopeptides reported in the APD database was only 37.5%, and it was inferred that it was a novel antimicrobial peptide. Surfactin and the hexapeptide showed antibacterial activity against both Gram-positive and Gram-negative bacteria.

Key words: Bacillus amyloliquefaciens; antibacterial activity; isolation; purification; antibacterial peptides

中图分类号:

Q939

姚佳明，田亚平. 解淀粉芽孢杆菌抑菌肽的分离鉴定及其抑菌谱表征[J]. 食品科学, 2020, 41(16): 126-131.

YAO Jiaming, TIAN Yaping. Isolation and Identification of Antimicrobial Peptides Produced by Bacillus amyloliquefaciens and Characterization of Their Antibacterial Spectra[J]. FOOD SCIENCE, 2020, 41(16): 126-131.

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解淀粉芽孢杆菌抑菌肽的分离鉴定及其抑菌谱表征

Isolation and Identification of Antimicrobial Peptides Produced by Bacillus amyloliquefaciens and Characterization of Their Antibacterial Spectra

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