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Characterization of contaminating bacterial community from ethanol fermentation process and the impact on the metabolome

Full text
Author(s):
Maria Letícia Bonatelli
Total Authors: 1
Document type: Doctoral Thesis
Press: Piracicaba.
Institution: Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALA/BC)
Defense date:
Examining board members:
Carlos Alberto Labate; Welington Luiz de Araujo; João Lucio de Azevedo; Andreas Karoly Gombert; Carlos Eduardo Vaz Rossell
Advisor: Carlos Alberto Labate
Abstract

The fermentation of Saccharomyces cerevisiae for ethanol production has great importance to Brazil since it is responsible for the production of a renewable energy source that is widely used in the automotive industry. However, in industrial scale, the yeast fermentation does not occur in an aseptic environment, where different contaminant microorganisms are capable of growing, competing for nutrients and even interfering with the S. cerevisiae fermentation. In order to better understand who the contaminating microorganisms are and what they do in the fermenter, it was used one polyphasic approach. The survey of the bacterial microflora present in two different São Paulo state distilleries was carried out by cultivation independent techniques. Subsequently, fermentation assays were performed with S. cerevisiae CAT-1 in the presence of the contaminant Lactobacillus fermentum (I-2) to analyze the interaction of these microorganisms in the fermenter. The analysis was performed through metabolomics accessed by gas chromatography-mass spectrometry (GC-MS) and, for that, initially was established an efficient methodology for fermentation metabolite analysis by GC-MS. In the distilleries, it was reported a higher percentage of Lactobacillus than ever described and bacterial population appeared to be characteristic of each distillery and persistent over time. After the establishment of fermentation metabolite analysis methodology by GC-MS, it was possible to identify 261 metabolites, and the three most abundant classes were Carbohydrates and carbohydrate conjugates (16%), Carboxylic acids and derivatives (12%) and Fatty acyls ( 5%). In the fermentation assay of S. cerevisiae CAT-1 in the presence of the contaminant L. fermentum (I-2), it was possible to identify 208 metabolites, where 50 were differentially abundant. In addition, the glycolytic pathway was enhanced in the fermentation of S. cerevisiae CAT-1, and in fermentation of S. cerevisiae (CAT-1) in the presence of L. fermentum (I-2), the production of amino acids from glutamate appeared to be important. Thus, a polyphasic analysis can help in the understanding of the relationship between contaminating microorganisms with the yeast in the fermenter. (AU)

FAPESP's process: 13/08431-0 - Microbial diversity presented on fermented juice from burned and mechanized harvesting of sugarcane and the impact on the fermentation metabolome
Grantee:Maria Leticia Bonatelli
Support Opportunities: Scholarships in Brazil - Doctorate