Protein extraction method for the proteomic study of a Mexican traditional fermented starchy food
Por:
Cárdenas C., Barkla B.J., Wacher C., Delgado-Olivares L., Rodríguez-Sanoja R.
Publicada:
5 dic 2014
Resumen:
Pozol is a traditional fermented maize dough prepared in southeastern
Mexico. Wide varieties of microorganisms have already been isolated from
this spontaneously fermented product; and include fungi, yeasts, and
lactic- and non-lactic acid bacteria. Pozol presents physicochemical
features different from that of other food fermentation products, such
as a high starch content, in addition to a low protein content. It is
these qualities that make it intractable for protein recovery and
characterization. The aim of this study was to develop a methodology to
optimize the recovery of proteins from the pozol dough following
fermentation, by reducing the complexity of the mixture prior to 2D-PAGE
analysis and sequencing, to allow the characterization of the
metaproteome of the dough. The proteome of 15 day fermented maize dough
was characterized; proteins were separated and analyzed by mass
spectrometry (LC MS/MS). Subsequent sequence homology database
searching, identified numerous bacterial and fungi proteins; with a
predominance of lactic acid bacterial proteins, mainly from the
Lactobacillus genus. Fungi are mainly represented by Aspergillus. For
dominant genera, the most prevalent proteins belong to carbohydrate
metabolism and energy production, which suggest that at 15 days of
fermentation not only fungi but also bacteria are metabolically active.
Biological significance
Several methodologies have been employed to study pozol, with a specific
focus toward the identification of the microbiota of this fermented
maize dough, using both traditional cultivation techniques and culture
independent molecular techniques. However to date, the dynamics of this
complex fermentation is not well understood. With the purpose to gain
further insight into the nature of the fermentation, we used proteomic
technologies to identify the origin of proteins and enzymes that
facilitate substrate utilization and ultimately the development of the
microbiota and fermentation. In this paper we overcome the first general
challenge for such studies, obtaining a protein sample with adequate
quality capable of representing the system. (C) 2014 Elsevier B.V. All
rights reserved.
Filiaciones:
Cárdenas C.:
Univ Nacl Autonoma Mexico, Programa Posgrado Ciencias Bioquim, Mexico City 04510, DF, Mexico
Univ Nacl Autonoma Mexico, Inst Invest Biomed, Mexico City 04510, DF, Mexico
Barkla B.J.:
Univ Nacl Autonoma Mexico, Inst Biotechnol, Cuernavaca 62210, Morelos, Mexico
Wacher C.:
Univ Nacl Autonoma Mexico, Fac Quim, Mexico City 04510, DF, Mexico
Delgado-Olivares L.:
Facultad de Química, Universidad Nacional Autónoma de México, Distrito Federal, 04510, Mexico
Rodríguez-Sanoja R.:
Univ Nacl Autonoma Mexico, Inst Invest Biomed, Mexico City 04510, DF, Mexico
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