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链霉菌利用稻草作为发酵底物生产去甲万古霉素抗生素。

Production of destomycin-A antibiotic by Streptomyces sp. using rice straw as fermented substrate.

作者信息

Atta H M, Abul-Hamd A T, Radwan H G

机构信息

Botany and Microbiology Dept., Faculty of Science (Boys), Al-Azhar University, Cairo, Egypt.

出版信息

Commun Agric Appl Biol Sci. 2009;74(3):879-97.

Abstract

Hundred and twenty microbial isolates could be isolated from different soil samples collected from different localities in Egypt. One of the actinomycete culture AZ-H-A5 from three cultures was found to produce a wide spectrum antimicrobial agent when cultivated on rice straw. The actinomycete AZ-H-A5 could be isolated from a soil sample collected from Helwan district, Egypt. The nucleotide sequence of the 16s RNA gene (1.5 Kb) of the most potent strain evidenced an 85% similarity with Streptomyces pseudovenezue, EU841712 and Streptomyces galilaeus. From the taxonomic features, the actinomycetes isolate AZ-H-A5 matches with Streptomyces rimosus in the morphological, physiological and biochemical characters. Thus, it was given the suggested name Streptomyces rimosus, AZ-H-A5. The parameters controlling the biosynthetic process of antimicrobial agent formation including: inoculum size, different pH values, different temperatures, different incubation period, and different carbon and nitrogen sources, potassium nitrate, K2HPO4, MgSO4.7H2O and KCl concentrations were fully investigates. The active metabolite was extracted using ethyl acetate (1:1, v/v) at pH 7.0. The separation of the active ingredient and its purification was performed using both thin layer chromatography (TLC) and column chromatography (CC) techniques. The physicochemical characteristics of the purified antibiotic viz. color, melting point, solubility, elemental analysis, spectroscopic characteristics and chemical reactions have been investigated. This analysis indicates a suggested empirical formula of C20H37N13O13. The minimum inhibition concentrations "MICs" of the purified antimicrobial agent were also determined. The purified antimicrobial agent was suggestive of being belonging to Destomycin-A antibiotic produced by Streptomyces rimosus, AZ-H-A5.

摘要

从埃及不同地区采集的不同土壤样本中分离出了120株微生物菌株。在三种培养物中,其中一种放线菌培养物AZ-H-A5在稻草上培养时能产生广谱抗菌剂。放线菌AZ-H-A5可从埃及赫尔万地区采集的土壤样本中分离得到。最具活性菌株的16s RNA基因(1.5 Kb)的核苷酸序列与委内瑞拉链霉菌(Streptomyces pseudovenezue)EU841712和加利利链霉菌(Streptomyces galilaeus)有85%的相似性。从分类学特征来看,放线菌分离株AZ-H-A5在形态、生理和生化特征上与龟裂链霉菌(Streptomyces rimosus)匹配。因此,它被赋予了建议名称龟裂链霉菌AZ-H-A5。对抗菌剂形成生物合成过程的控制参数进行了全面研究,包括接种量、不同的pH值、不同的温度、不同的培养时间以及不同的碳源和氮源、硝酸钾、磷酸氢二钾、七水硫酸镁和氯化钾浓度。在pH 7.0时用乙酸乙酯(1:1,v/v)提取活性代谢物。使用薄层色谱(TLC)和柱色谱(CC)技术对活性成分进行分离和纯化。对纯化抗生素的物理化学特性,即颜色、熔点、溶解度、元素分析、光谱特征和化学反应进行了研究。该分析表明其经验式为C20H37N13O13。还测定了纯化抗菌剂的最低抑菌浓度(MICs)。纯化的抗菌剂提示属于龟裂链霉菌AZ-H-A5产生的 Destomycin-A抗生素。

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