银杏种仁酚酸的纯化、鉴定及其抑菌活性分析  

Purification, Identification and the Antimicrobial Activity of Ginkgolic Acids in Ginkgo Seeds

作　　者：吴海霞[1,2] 吴彩娥[1] 刘金达[1,3] 范龚健[1] 李婷婷[1] 应瑞峰[1] 

机构地区：[1]南京林业大学轻工科学与工程学院,南京210037 [2]运城学院生命科学系,山西运城044000 [3]沂水县林业局,山东临沂276400

出　　处：《中国食品学报》2015年第3期207-215,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：江苏省普通高校研究生科研创新计划项目(CXZZ12_0544);江苏省科技支撑(农业)项目(BE2011381)

摘　　要：以银杏种仁为材料,采用乙醇浸提、石油醚萃取的方法提取酚酸,研究银杏种仁酚酸硅胶柱层析纯化工艺。采用液相色谱(HPLC)及液-质联用(LC-MS)分析银杏种仁中酚酸的组成及含量。以2种细菌及4种真菌为供试菌,比较银杏种仁酚酸的抑菌活性,并确定其对各菌种的最小抑菌浓度(MIC)。结果表明,以流速为1.2 m L/min的石油醚-乙醚-甲醇(89∶11∶1,V/V/V)为洗脱液,对银杏酚酸粗提物进行硅胶柱层析纯化,取得较好的纯化效果。HPLC及LC-MS分析表明:银杏酚酸主要由C13∶0,C15∶1,C17∶2,C15∶0和C17∶1等5种酚酸组分组成。以C13∶0,C15∶1,C17∶1计,银杏种仁中的酚酸含量为0.11 g/kg脱脂干粉,其中C13∶0,C15∶1,C17∶1的含量分别为0.013 g/kg脱脂干粉、0.042 g/kg脱脂干粉、0.055 g/kg脱脂干粉。抑菌试验表明,银杏酚酸对供试的2种细菌(大肠杆菌和枯草芽孢杆菌)以及4种真菌(青霉、产紫青霉、沙门柏干酪青霉、黑曲霉)有抑制作用。银杏酚酸对大肠杆菌、枯草芽孢杆菌及青霉菌有较好的抑菌效果,其MIC分别为7.5,15和25 mg/m L。Objective： To purify and identify quorum sensing signal peptide of Lactobacillus pentosus 31-1. Methods：The active principle was purified to homogeneity by ammonium sulfate precipitation, ultrafilteration, hydrophobic interaction chromatography, gel filtration chromatography and reverse phase high-performance liquid chromatography（RP-HPLC）.MALDI-TOF mass spectrometer was used for determination molecular weight of the purified peptide AIP, and its N-terminal sequences was analyzed by Edman degradation. Closantel, a histidine protein kidnase inhibitor was used to verify the role of the purified peptide. Results： The purification resulted in an 800-fold increase in specific activity with a yield of 160 000 IU/m L of the induction activity. The mass of the purified peptide AIP was 2 985.16 u, its partial N-terminal sequence was： NH2-KSSAYSLQMGATAIK. Bacteriocin production inhibited by closantel could not be restored by peptide AIP, which further confirmed its quorum sensing signal molecule role. Conclusion： The obtaining of the quorum sensing peptide AIP laid the fundation for the regulation of bacteriocin fermentation in L. pentosus 31-1.

关 键 词：银杏种仁 酚酸 纯化 高效液相色谱 液质联用 抑菌活性 

分 类 号：O657[理学—分析化学] TS255.6[理学—化学]