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小链霉菌产去铁胺E的优化

Optimization of desferrioxamine E production by Streptomyces parvulus.

作者信息

Gáll Tamás, Lehoczki Gábor, Gyémánt Gyöngyi, Emri Tamás, Szigeti Zsuzsa M, Balla György, Balla József, Pócsi István

机构信息

1 MTA-DE Vascular Biology, Thrombosis and Hemostasis Research Group, Hungarian Academy of Sciences , Debrecen, Hungary.

2 Department of Inorganic and Analytical Chemistry, Faculty of Science and Technology, University of Debrecen , Debrecen, Hungary.

出版信息

Acta Microbiol Immunol Hung. 2016 Dec;63(4):475-489. doi: 10.1556/030.63.2016.029.

Abstract

Siderophores are produced by a number of microbes to capture iron with outstandingly high affinity, which property also generates biomedical and industrial interests. Desferrioxamine E (DFO-E) secreted by streptomycetes bacteria can be an ideal candidate for iron chelation therapy, which necessitates its cost-effective production for in vitro and animal studies. This study focused on the optimization of DFO-E production by Streptomyces parvulus CBS548.68. Different combinations of various carbon and nitrogen sources as well as the addition of 3-morpholinopropane-1-sulfonic acid (MOPS) markedly affected DFO-E yields, which were attributed, at least in part, to the higher biomass productions found in MOPS-supplemented cultures. In MOPS-supplemented glucose and sodium glutamate medium, DFO-E productions as high as 2,009 ± 90 mg/l of culture medium were reached. High-performance liquid chromatography analysis demonstrated that a simple two-step purification process yielded DFO-E preparations with purities of ∼97%. Matrix assisted laser desorption ionization-time of flight mass spectrometry analysis showed that purified DFO-E always contained traces of desferrioxamine D2.

摘要

许多微生物会产生铁载体,以极高的亲和力捕获铁,这一特性也引发了生物医学和工业领域的兴趣。链霉菌分泌的去铁胺E(DFO-E)可能是铁螯合疗法的理想候选药物,因此需要以具有成本效益的方式生产,用于体外和动物研究。本研究聚焦于优化小链霉菌CBS548.68生产DFO-E的工艺。各种碳源和氮源的不同组合以及添加3-吗啉丙烷-1-磺酸(MOPS)对DFO-E的产量有显著影响,这至少部分归因于在添加MOPS的培养物中发现的更高生物量产量。在添加MOPS的葡萄糖和谷氨酸钠培养基中,DFO-E的产量高达2009±90毫克/升培养基。高效液相色谱分析表明,简单的两步纯化过程可得到纯度约为97%的DFO-E制剂。基质辅助激光解吸电离飞行时间质谱分析表明,纯化后的DFO-E总是含有痕量的去铁胺D2。

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