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Analysis of the Biosynthesis Conditions and Characteristics of Siderophores from Pseudomonas sp. SPF-1

Author: ChenShaoXing
Tutor: XieZhiXiong
School: Wuhan University
Course: Microbiology
Keywords: Siderophore Pseudomonas Iron ions Spectral absorption CAS detection
CLC: Q936
Type: Master's thesis
Year: 2005
Downloads: 321
Quote: 7
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Abstract


In this paper, CAS testing methods Pseudomonas sp.SPF-1 strain of the basic characteristics of siderophore production and siderophore circle or by measuring fluorescence colony diameter circle diameter ratio, UV - visible continuous wavelength scanning method, △ OD 630 , OD 600 , OD 400 absorption methods such as Pseudomonas sp.SPF-1 strains produce siderophores conditions, iron ions on its ability to produce siderophores effects were studied, and the separation and purification of Pseudomonas sp.SPF-1 strains siderophores and a preliminary analysis of its properties. The results showed that: Pseudomonas sp.SPF-1 strain has a generic CAS tablet faster, stronger production circle characteristic; siderophore producing optimum temperature of 28-30 ℃, the optimum pH of 7.5-8.0, and more adequate ventilation, the greater the amount siderophore secretion; through with the other seven siderophore producing Pseudomonas comparison found that different strains of Pseudomonas there are big differences, Pseudomonas sp.SPF-1 strains produce siderophores particularly in the production capacity of fluorescent siderophore strong; culture medium by a comparative study found that sucrose - aspartic acid (MSA) culture medium was more sensitive than the generic CAS, particularly suitable for the detection of fluorescent siderophore. Iron ions than 1μM Pseudomonas sp.SPF-1 can promote the growth of strains of iron, but severely inhibit the secretion vector, which results in a liquid culture medium is more obvious than the solid culture medium; while 2-4μM of iron chelator can be promote the secretion of siderophores, and divalent cation chelator no effect; and found fluorescent and non-fluorescent siderophore iron on iron ion response differences, and the fluorescent siderophore iron ions in response aspects of siderophore sensitive than non-fluorescent; using Sephadex G-25 对 Pseudomonas sp.SPF-1 were isolated and purified siderophore preliminary nature of the study: it is water-soluble secreted siderophores, with high thermal stability; in the 400nm absorption peak at a specific, pH, affect its maximum absorption wavelength; fluoresces under ultraviolet light, fluorescence and iron ions extracted off, and the absorption peak at 400nm remains; purified siderophore Pseudomonas sp.SPF-1 with the same CAS detection in general formed on the iron plate Carrier halo; by agarose gel electrophoresis and thin layer chromatography found Pseudomonas sp.SPF-1 secretion in three kinds of fluorescent siderophores, and the polarity of the large differences; wavelength scan findings with fluorescein in the spectral absorption (490nm), iron ion transport essential differences exist. Research results show that we have established MSA-CAS is a highly sensitive detection method, particularly suitable for fluorescence detection method siderophore; discovery fluorescent and non-fluorescent siderophores in iron ions acknowledge differences, and the former than the latter sensitive phenomenon, This is discussed in more depth Pseudomonas siderophore synthesis and provide the basis for regulatory mechanisms.

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