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13C, and heteronuclear quantum coherence NMR.
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note
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Surface tension measurements were made with a Fisher Model 21 Tensiomat with a 5.99-IR loop and jacketed 20-ml cell thermostated at 25°C. The CMC value was estimated from the intersection of the extrapolated linear portions of the plot of surface tension versus siderophore concentration.
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26
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0343307709
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note
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The CMC of desferri-marinobactin E at pH 3, at which the siderophore has no net charge, is 25 μM. Under these conditions, the CMC does not vary with ionic strength (for 0 to 1.0 M KCl). However, at pH 7.0, at which the head group bears a - 1 charge due to deprotonation of the terminal carboxylate group, the related desferri-marinobactin D has an ionic strength-dependent CMC, ranging from 100 μM at 0 M KCl to 35 μM at 1.0 M KCl.
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27
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0004130696
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VCH, New York
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Dynamic light-scattering measurements were performed on Fe(III)-marinobactin vesicles prepared by addition of ferric nitrate to a solution of the desferri form of the marinobactins in 100 mM tris-HCl (pH 8.0). Scattered light from a 30-mW, 633-nm helium-neon laser at an angle of 90° was collected and analyzed with a Brookhaven Instruments BI-9000AT Digital Correlator to determine the effective diffusion constant of the vesicles, which was converted to hydrodynamic radius [D. F. Evans and H. Wennerstrom, The Colloidal Domain (VCH, New York, 1994)].
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16S ribosomal RNA (rRNA) sequences of marine siderophore producers (accession numbers AF199438, AF199439, and AF199440) were obtained from polymerase chain reaction products with standard primers (20). Both strands were sequenced, and the final sequences were aligned to the E. coli secondary structure (21). For phylogenetic analysis, sequences were obtained from the Ribosomal Database Project (RDP) (22) and GenBank and aligned with Sequencher. Accession numbers of sequences used for the analysis in Fig. 2 are M11223, U14584, M93352, L42619, M93358, M93357, L42618, X71814, AJ000726, AF148811, X67022, U85854, AJ000647, AB006769, U26262, X76334, and X74697. Oceanospirillum maris is an RDP sequence. Hypervariable regions were excluded, and a final data set of 1327 characters was analyzed with maximum parsimony in PAUP version 4.02b (23) with a transversion cost of two. Bootstrapping was done 1000 times with a heuristic search algorithm with 10 rounds of random addition at each replication. H. aquamarina and H. meridiana have indistinguishable 16S rRNA sequences, and the species status of H. meridiana is questionable. We compared phenotypic characters of the type strains of both species and strain DS40M3 and found the differences to be insignificant. Despite slight sequence differences between H. aquamarine and DS40M3, it is most conservative to regard it as a strain
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