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Metabolites. 2014 Apr 22;4(2):218-31. doi: 10.3390/metabo4020218.

Xanthan Gum Removal for 1H-NMR Analysis of the Intracellular Metabolome of the Bacteria Xanthomonas axonopodis pv. citri 306.

Author information

1
Brazilian Biosciences National Laboratory, Rua Giuseppe Máximo Scolfaro, 10.000-Polo II de Alta Tecnologia, Caixa Postal 6192, CEP 13083-970, Campinas/SP, Brazil. vanessa.pegos@lnbio.cnpem.br.
2
Brazilian Biosciences National Laboratory, Rua Giuseppe Máximo Scolfaro, 10.000-Polo II de Alta Tecnologia, Caixa Postal 6192, CEP 13083-970, Campinas/SP, Brazil. rafael.canevarolo@lnbio.cnpem.br.
3
Brazilian Biosciences National Laboratory, Rua Giuseppe Máximo Scolfaro, 10.000-Polo II de Alta Tecnologia, Caixa Postal 6192, CEP 13083-970, Campinas/SP, Brazil. aline.sampaio@lnbio.cnpem.br.
4
Brazilian Biosciences National Laboratory, Rua Giuseppe Máximo Scolfaro, 10.000-Polo II de Alta Tecnologia, Caixa Postal 6192, CEP 13083-970, Campinas/SP, Brazil. andrea.balan@lnbio.cnpem.br.
5
Brazilian Biosciences National Laboratory, Rua Giuseppe Máximo Scolfaro, 10.000-Polo II de Alta Tecnologia, Caixa Postal 6192, CEP 13083-970, Campinas/SP, Brazil. ana.zeri@lnbio.cnpem.br.

Abstract

Xanthomonas is a genus of phytopathogenic bacteria, which produces a slimy, polysaccharide matrix known as xanthan gum, which involves, protects and helps the bacteria during host colonization. Although broadly used as a stabilizer and thickener in the cosmetic and food industries, xanthan gum can be a troubling artifact in molecular investigations due to its rheological properties. In particular, a cross-reaction between reference compounds and the xanthan gum could compromise metabolic quantification by NMR spectroscopy. Aiming at an efficient gum extraction protocol, for a 1H-NMR-based metabolic profiling study of Xanthomonas, we tested four different interventions on the broadly used methanol-chloroform extraction protocol for the intracellular metabolic contents observation. Lower limits for bacterial pellet volumes for extraction were also probed, and a strategy is illustrated with an initial analysis of X. citri's metabolism by 1H-NMR spectroscopy.

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