摘要
为从海洋微生物中获取天然α-葡萄糖苷酶抑制活性(α-glucosidase inhibiting,α-GI)化合物,对一株前期研究发现具有α-GI活性的细菌进行鉴定和培养条件优化,并对其代谢产物进行分离,获取和鉴定其活性化合物。通过形态学观察和16S r DNA测序鉴定活性菌株HY95为波茨坦短芽孢杆菌(Brevibacillus borstelensis);采用分析单因素和正交试验选取菌株的最佳培养条件为:2.5%(V/V)的接种量,130 r/min的摇床转速,28℃恒温培养60 h。经优化后的MB培养基中:蛋白胨5.00 g/L,酵母粉1.50g/L、氯化钠9.725 g/L,pH 7.5;以生物活性测试为导向,用化学方法(薄层色谱、高效液相色谱)对其中的活性组分进行分离纯化,并经核磁共振氢谱分析确定得到一个α-GI活性化合为环(苯丙氨酸-酪氨酸),其对α-葡萄糖苷酶的抑制率为53.72%±4.92%。为α-GI活性化合物的筛选提供了一个极有开发前景的途径和来源。
Aiming at isolating naturalα-glucosidase inhibiting （α-GI） compounds from marine microorganisms, this paper identified anα-GI active bacterium （HY95） which was screened by previous study. Its culture conditions were optimized and theα-GI active compounds were isolated from it. Through the morphological observation and the analysis of 16S rDNA sequence, the active strain of HY95 was identified asBrevibacillus borstelensis.By using single factor analysis and orthogonal test, the culture conditions were optimized as inoculum ratio of 2.5% （V/V）, shaker speed of 130 rpm, culture temperature of 28℃, and incubation time of 60 h in the modified marine broth medium with 5.00 g/L peptone, 1.50 g/L yeast extract, 9.725 g/L sodium chloride, and the initial pH of 7.5. The active compounds were separated using the column chromatography, thin layer chromatography （TLC）, and high performance liquid chromatography （HPLC） with the guidance of bioactive test. The final purified compound was subjected to H1 nuclear magnetic resonance （1H NMR） and was identified as the Cyclo（Tyr-Phe）. Theα-GI activity of the pure compound was verified with inhibition rate of 53.72±4.92%. The results of the present study provide a promising direction and source for efficientα-GI compounds.
出处
《海洋科学》
CAS
CSCD
北大核心
2015年第10期36-44,共9页
Marine Sciences
基金
国家自然科学基金资助项目(41076097
41106113
41271521)
教育部科学技术研究重点项目资助项目(211065)
江苏省自然科学基金(BK2012267)
江苏省教育厅高校自然科学基金资助项目(11KJB170011)
关键词
α-葡萄糖苷酶抑制活性
菌株鉴定
培养条件优化
活性化合物分离
α-glucosidase inhibiting activity
strain identification
optimization of culture conditions
isolation of active compounds
