Supporting Information. The TLC analysis of the carbomycobactin (silica gel) column fractions and the LC-MS data for column fraction# 11, which contained the carbomycobactins used in this study.

Supporting Information

Utilization of Fe+3-Acinetoferrin Analogues as an iron source by Mycobacterium tuberculosis

G. Marcela Rodriguez*, Richard Gardner,† Navneet Kaur,† and Otto Phanstiel IV†

*communicating author, The Public Health Research Institute at the International Center for Public Health, University of Medicine and Dentistry of New Jersey, 225 Warren Street, Newark, NJ 07103-3535,

† Department of Chemistry, University of Central Florida, Orlando, FL 32816-2366

Table of Contents

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TLC Analysis of carboxymycobactin fractions S2-S3

LC-MS data for fraction 11 S4-S10

(Note: LC-MS revealed a homologous series of carboxymycobactins (2) were present in fraction 11, an average MW of these homologues was estimated as 785 g/mol. This MW was used for generating solutions of 2 used in the manuscript).


TLC Analysis. TLC plate analysis of the carboxymycobactin column chromatography fractions (5% MeOH in CHCl3; fractions #1-18). Bands were observed by illuminating the TLC plate with short wavelength UV light.TLC conditions eluted the material using 5%MeOH in CHCl3 on a ‘normal-phase’ silica gel TLC plate. The far right lane is the crude extract (“C”, as a control to show the ladder bands of the crude extract) and from far left to right are fractions #1-18. Of these fractions, fraction 11 was shown by LC-MS to contain the carboxymycobactin as a mixture of homologues differing by incremental CH2 groups (DMW = 14; see the LC-MS report in this Supporting Information).Note: Fractions 6 and 7, which look dark in the image, did not contain material in the proper MW range.

Image of TLC plate (column fract #s are listed both on the image and below to assist the reader).

fract# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Crude

Note: other authors have isolated exochelins from mycobacteria using similar methods. Macham, L.P.; Ratledge, C.; Nocton, J.C. Extracellular Iron Acquisition by Mycobacteria: Role of the Exochelins and Evidence Against the Participation of Mycobactin. Infection and Immunity, 1975, 12, No. 6, 1242-1251.


Figure 1. Fraction 11, (+)ESI-MS mass chromatograms of the [M+H]+ ions associated with the major UV peaks. Use of these areas to calculate

the average MW resulted in an average MW = 785 g/mol for carboxymycobactin, 2


Figure 2. MW 772.



Figure 3. MW 770

Figure 4. MW 786.



Figure 5. MW 784.



Figure 6. MW 798.



Figure 7. MW 800: MSn spectra contain ions form MW 798 and MW 800.

S 1