昆虫学报 ›› 2023, Vol. 66 ›› Issue (10): 1282-1288.doi: 10.16380/j.kcxb.2023.10.002

• 研究论文 • 上一篇    下一篇

黑翅土白蚁肠道放线菌菌株BYC-18及其抗菌代谢产物的分离鉴定

尹彩萍#,*, 白雪妍#, Naeem ABBAS, 孙璐璐, 殷欣冉, 张应烙   

  1. (安徽农业大学生命科学学院, 合肥 230036)
  • 出版日期:2023-10-20 发布日期:2023-11-27

Isolation and identification of actinomycete strain BYC-18 and its antimicrobial metabolites from the gut of Odontotermes formosanus (Isoptera: Termitidae)

YIN Cai-Ping#,*, BAI Xue-Yan#, Naeem ABBAS, SUN Lu-Lu, YIN Xin-Ran, ZHANG Ying-Lao   

  1.  (College of Life Sciences, Anhui Agricultural University, Hefei 230036, China)
  • Online:2023-10-20 Published:2023-11-27

摘要:

【目的】测定黑翅土白蚁肠道放线菌发酵产物的抗菌活性,并对其抗菌活性成分进行分析, 以发现新颖的抗菌先导化合物。【方法】采用涂布平板法对黑翅土白蚁肠道放线菌进行分离;通过牛津杯法测试菌株发酵液提取物对4种致病菌(金黄色葡萄球菌Staphylococcus aureus、大肠杆菌Escherichia coli、枯草芽孢杆菌Bacillus subtilis和白色念珠菌Candida albicans)的抗菌活性,筛选出活性菌株BYC-18;通过形态学特征和16S rRNA序列分析确定BYC-18的分类学地位;采用滤纸片法测定BYC-18发酵液在不同极性溶剂萃取物的抗菌活性;运用多种色谱方法从乙酸乙酯粗提物中分离纯化抗菌活性化合物,利用质谱和核磁共振谱鉴定其化学结构;采用滤纸片法和最低抑制浓度法测定分离的化合物的抗菌活性。【结果】BYC-18被鉴定为链霉菌属Streptomyces sp.菌株,该菌发酵液对4种致病菌均有抗菌活性且其乙酸乙酯萃取物对金黄色葡萄球菌的抑制作用明显,抑菌圈直径达11.1 mm。从乙酸乙酯萃取物中分离得到1个单体化合物BYC-18-1,经鉴定为β-玉红霉素(β-rubromycin);抗菌活性测试表明,在供试浓度为30 μg/6 mm滤纸片时,β-玉红霉素对金黄色葡萄球菌具有较强的抑制作用,抑菌圈直径为13.2 mm,与阳性对照硫酸庆大霉素的抑菌活性(抑菌圈直径: 16.6 mm)接近。【结论】链霉菌菌株BYC-18具有开发为新型微生物源杀菌剂的潜力。

关键词: 黑翅土白蚁, 肠道放线菌, 链霉菌, 抗菌活性, 天然产物

Abstract: 【Aim】To determine the antimicrobial activities of the fermentation products of actinomycetes from the gut of Odontotermes formosanus and analyze their antimicrobial components, so as to find novel antimicrobial lead compounds.【Methods】 The gut actinomycetes of O. formosanus were isolated by spread plate method. The antimicrobial activities of the fermentation broth extracts against four pathogens (Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Candida albicans) were tested by the Oxford cup method, and the active strain BYC-18 was screened. The taxonomic status of BYC-18 was determined according to its morphological features and 16S rRNA sequence analysis. The antimicrobial activities of different polar solvent extracts of BYC-18 fermentation solution were determined by filter paper method. Active compounds were isolated and purified from ethyl acetate crude extract by various chromatographic methods, and the chemical structure of the isolated compound was identified by mass spectrometry and nuclear magnetic resonance spectroscopy. The antimicrobial activities of the isolated compound were determined by filter paper method and minimum inhibitory concentration method. 【Results】 BYC-18 was identified as a strain of Streptomyces sp. The fermentation broth of BYC-18 had antimicrobial activities against all the four pathogens, and its ethyl acetate extract had an obvious inhibitory effect on S. aureus, with an inhibition zone diameter of 11.1 mm. A monomer compound BYC-18-1 was isolated from the ethyl acetate extract and identified as β-rubromycin. The antimicrobial activity test showed that β-rubromycin had a strong inhibitory effect on S. aureus at the test concentration of 30 μg/6 mm filter paper, and its inhibition zone diameter was 13.2 mm, which was close to that of the positive gentamicin sulfate (inhibition zone diameter: 16.6 mm). 【Conclusion】The Streptomyces strain BYC-18 has the potential to be developed as a new bactericide.

Key words: Odontotermes formosanus, gut actinomycetes; Streptomyces sp., antimicrobial activity, natural product