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链霉菌702菌株所产抗真菌单体组分DZP-8和DZP-9的分离纯化与制备
  • ISSN号:1004-874X
  • 期刊名称:《广东农业科学》
  • 时间:0
  • 分类:S482.28[农业科学—农药学;农业科学—植物保护]
  • 作者机构:[1]江西农业大学生物科学与工程学院,江西南昌330045
  • 相关基金:国家自然科学基金(30960011,31071724); 江西省自然科学基金(2009GZN0030); 江西省科技支撑计划项目(2007BN14002)
中文摘要:

用乙醇浸提链霉菌702菌株发酵液,滤液真空浓缩成油状物,再用乙酸乙酯萃取,萃取液真空浓缩成浸膏,浸膏通过硅胶柱层析,以氯仿与甲醇不同配比进行梯度洗脱,收集含抗真菌活性物质馏分洗脱液,进行真空浓缩至干燥为抗真菌活性物质粗品。粗品通过制备型HPLC仪进一步分离纯化,用乙腈∶水∶TFA不同浓度进行梯度洗脱,按单组分保留时间收集不同馏分洗脱液,将洗脱液冻干成抗真菌活性单体组分样品。抗菌活性和单组分样品纯度检测分别用管碟法检测对黑曲霉的抑菌活性和分析型HPLC。结果表明,含抗真菌活性单位10 000 mg/L发酵液200 L经乙醇浸提液浓缩后得到含抗真菌活性单位22 500 mg/L的油状物80 L,浸提率为90%;80 L油状物经乙酸乙酯萃取浓缩得含抗真菌活性单位120 000 mg/L的12 L浸膏,萃提率为80%。500 g浸膏经硅胶柱层析,获得含抗真菌活性单位达750 mg/g的粗品48.0802 g。粗品经制备型HPLC仪获得纯度分别达到99.47%和90.28%的抗真菌单组分DZP-8和DZP-9。

英文摘要:

From a large-scale fermentation of Streptomyces spp.702,mycelium was extracted from the culture filtrate.Filtration was concentrated under reduced pressure to a brown gum.The brown gum was partitioned in AcOEt.The combined extracts were concentrated under reduced pressure.The AcOEt extract was mixed on silica gel and fractionated through flash chromatography eluting with a gradient mixture of chloroform-methanol giving some fractions of antifungal activity.The crude extract was further separated on preparing high pressure liquid chromatography system with mobile phase acetonitrile,water and TFA gradient elution.Two antifungal compounds,which had inhibitory activity against various fungal pathogens were purified.The antimicrobial activity of the fractions was analyzed by cup-plate method on Aspergillusniger,which determined purity of compound by analyzing HPLC.From a large-scale fermentation(200 L,antifungal activity unit 10 000 mg/L),mycelium was extracted.Combined two filtrate to give 80 L of concentrated liquid(antifungal activity unit 22 500 mg/L).The extraction rates were 90%.After the ethanol extraction extrated by AcOEt and concentrated,a brown gum(12 L,antifungal activity unit 120 000 mg/L) was gained.The extraction rates were 80%.The combined extracts were concentrated under reduced pressure to yield 48.0802 g of a crude extract(antifungal activity unit 750 mg/g).Bioactivity-guided isolation by preparing HPLC provided two macrolactone DZP-8 and DZP-9.

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期刊信息
  • 《广东农业科学》
  • 北大核心期刊(2011版)
  • 主管单位:广东省农业科学院
  • 主办单位:广东省农业科学院 华南农业大学
  • 主编:
  • 地址:广州市天河区金颖路31号五山广东省农科院农经所
  • 邮编:510640
  • 邮箱:gdnykx@vip.163.com
  • 电话:020-38319948 38319941 38319942 38319946
  • 国际标准刊号:ISSN:1004-874X
  • 国内统一刊号:ISSN:44-1267/S
  • 邮发代号:
  • 获奖情况:
  • 广东省第二、第三届优秀科技期刊,广东省首届十佳期刊,中国期刊方阵“双效”期刊,第四届全国农业优秀期刊一等奖,第五届全车优秀农业期刊鑫犁奖一等奖,首届全国CAD规范一等奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国动物学记录,中国北大核心期刊(2008版),中国北大核心期刊(2011版)
  • 被引量:33995