摘要
采用微生物发酵法获取天然产物,是一项比植物提取法更为经济的环境友好技术。为建立一种微生物生产苦参碱的方法,本文从陕北油田石油污染土壤中分离、筛选和鉴定了一种能合成苦参碱的枝顶孢属真菌。首先采用薄层层析法、气相色谱-质谱联用法等分别确定了在所获真菌菌株发酵产物中含有苦参碱物质;使用光学显微镜观察发现,该菌株菌落呈白色绒毛状,呈圆形,菌丝分散,背面奶油色至赭黄色;菌丝无色,光滑且具隔膜,孢子囊梗直接从菌丝上长出,呈锥形,多单生;分生孢子无色,光滑,形成簇状或链状,长纺锤形,两头尖,未见厚垣孢子。内转录区间序列(internal transcribed sequence,ITS)扩增、测序后进行同源性比较发现,该菌株ITS序列与枝顶孢属的同源性高达99%。构建系统发育树表明,该真菌与枝顶孢属菌的遗传距离最近。综合形态学和遗传进化分析结果对此菌株进行了种属分类,确认这株能合成苦参碱物质的真菌应归属子囊菌门(Ascomycota)的枝顶孢属(Acremonium sp.)。
Matrine is one of the quinolizidine alkaloids.It is widely used as broad-spectrum agricultural pesticides,which exists mainly in roots of leguminous plants.Matrine has great prospects of research and development as it has very extensive pharmacological effects,mainly including analgesia,anti-inflammatory,anti-arrhythmic,antitumor,antiviral,immunosuppression and biological control.The raw material of obtaining matrine is mainly the traditional Chinese medicine sophora.But the extraction method of matrine from plants often has its drawbacks,such as the cumbersome extracting steps,the high extraction temperature,the complex operation process,and using plenty of organic solutions.In addition,the production efficiency is not high enough,and the product has many impurities.Also,this production process often results in a large number of loss in resource.As a consequence,the matrineproduction derived from plants has challenged with the requirements of industrial production,as it cannot meet the increasing market demand.The production of matrine by microbial fermentation,however,would be an alternative for the production obtained from plants,and must have more advantages over the extracting from plants,with less labor and time consumed.However,there are no many reports concerning microbes with capacity to produce matrine.Hence,it is very necessary to obtain matrine-producing microorganisms for establishing the bio-production by microbial fermentation.The study presented hereby was carried out to isolate and identify the fungus with the ability to synthesize matrine from petroleum-contaminated soil samples.The fungal isolate with the capability to synthesize matrine was classified by morphological and molecular biological methods.The fungal isolates were firstly obtained from samples of oil-contaminated soil in North Shaanxi,China,through spread-plate technique and plate streaking.The chemical compositions in fermentation broth of the obtained fungus were detected by thin-layer chromatography and gas chromatograph-mass spe
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2015年第5期586-592,共7页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
陕西省科技计划资助项目(693102)
中央高校科研业务费特别支持项目(1301030208)
关键词
苦参碱
石油污染
枝顶孢属
薄层层析
气相色谱-质谱联用
邻接法
matrine
petroleum-contaminated
Acremonium sp.
thin-layer chromatography
gas chromatographmass spectrometer
neighbor-joining method
