蕈状芽孢杆菌代谢产物增强小鼠免疫力活性物质的分离、鉴定  

Isolation and Identification of the Immunopotentiating Compound Prepared from the Metabolites of Bacillus Mycoides

作　　者：王高学[1] 高鸿涛[1] 付维法[1] 崔婧[1] 袁明[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《微生物学通报》2009年第6期858-864,共7页Microbiology China

基　　金：陕西省科技攻关项目(No2006K02-G14-03)

摘　　要：本文的目的是对蕈状芽孢杆菌代谢产物中增强小鼠免疫力的物质进行分离、鉴定和免疫增强作用测定。在生物活性追踪的指导下,本文通过大孔吸附树脂吸附、硅胶柱分离和葡聚糖凝胶柱纯化,对蕈状芽孢杆菌代谢产物中增强小鼠免疫力的活性成分进行了分离,得到了具有较强免疫增强作用的化合物(标记为M),并对化合物M进行了波谱测定,确定化合物M为环(脯氨酸-甘氨酸)二肽(C7H10O2N2)。以生理盐水为对照,通过腹腔注射,对小鼠进行了SOD活性、白细胞吞噬活性、白细胞杀菌活性3个指标测定。结果显示,在第14天时,SOD活性和白细胞吞噬活性达到了最高值,且同对照组相比有显著提高;在第21天时,白细胞杀菌活性达到了最高值,且同对照组相比有显著提高。以上结果表明,蕈状芽孢杆菌代谢产物中的环(脯氨酸-甘氨酸)二肽能够显著增强小鼠免疫力。An immunopotentiating compound has been isolated from the metabolites of Bacillus mycoides under the bioassay-guided isolation and identification for its immunopotentiating effect and chemical structure. The isolation and purification of the compound were consisted of macroporous adsorptive resins, silicagel chromatographic column and Sephadex G-200 chromatographic column. The immunopotentiating effect was assayed in every step isolation. At last, the only substance having the strongest immunopotentiating effect had been isolated and purified. Through the procedure consisiting of Ultra-Violet spectroscopy （UV）, IR （Infrared Radiation）, Time of Flight Mass Spectrum （TOF-MS）, Nuclear Magnetic Resonance （NMR） and Element analysis, the possible structure of compound M had been identified as cyclic （Pro-Gly） dipeptide （C7H10O2N2） （Diketopiperazine）. To be determined the immunopotentiating effect, the mice were treated by intraperitoneal injection of cyclic （Pro-Gly） dipeptide in the treatment group and distilled water in the control group. At the 14^th day after the injection, the SOD activity and the phagocytosis activities reached the peak value and were significantly higher than those in control group. At the 21^st day, the bactericidal activity reached peak value and was significantly higher than that in the control group. From the above results, we concluded that the main active component enhancing the immunity of mice was cyclic （Pro-Gly） dipeptide in the metabolism of Bacillus mycoides.

关 键 词：环(脯氨酸-甘氨酸)二肽 免疫增强剂 蕈状芽孢杆菌 分离纯化 鉴定 

分 类 号：Q93[生物学—微生物学]