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Investigating interspecific quorum sensing influence on cocoa fermentation quality through defined microbial cocktails

View ORCID ProfileO.G.G. Almeida, M. G. Pereira, R. L. Bighetti-Trevisan, E.S. Santos, E. G. De Campos, G.E. Felis, L.H.S. Guimarães, M.L.T.M Polizeli, B. S. De Martinis, View ORCID ProfileE.C.P. De Martinis
doi: https://doi.org/10.1101/2022.06.14.496151
O.G.G. Almeida
1Universidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas
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M. G. Pereira
2Universidade do Estado de Minas Gerais, Unidade Passos
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R. L. Bighetti-Trevisan
3Universidade de São Paulo, Faculdade de Odontologia de Ribeirão Preto. Departamento de Biologia Básica e Oral
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E.S. Santos
1Universidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas
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E. G. De Campos
4Appalachian State University, Department of Chemistry and Fermentation Sciences, Boone, NC, United States
7Universidade de São Paulo, Faculdade de Filosofia Ciências e Letras de Ribeirão Preto. Departamento de Química
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G.E. Felis
5University of Verona, Department of Biotechnology, Verona, Italy
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L.H.S. Guimarães
6Universidade de São Paulo, Faculdade de Filosofia Ciências e Letras de Ribeirão Preto. Departamento de Biologia
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M.L.T.M Polizeli
6Universidade de São Paulo, Faculdade de Filosofia Ciências e Letras de Ribeirão Preto. Departamento de Biologia
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B. S. De Martinis
7Universidade de São Paulo, Faculdade de Filosofia Ciências e Letras de Ribeirão Preto. Departamento de Química
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E.C.P. De Martinis
1Universidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas
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  • For correspondence: edemarti@usp.br
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Abstract

The fermentation of cocoa beans is a key process to supply high quality ingredients for the chocolate industry. In spite of several attempts to obtain standardised microbial cultures for cocoa fermentation, it is still a spontaneous process. It has been suggested lactobacilli present potential for quorum sensing (QS) regulation in cocoa fermentation, and in the present research, laboratory scale fermentations were carried out to further elucidate possible QS influence on microbial shifts and fermented seeds quality. The experimental design comprised the 96 hours-fermentations designated as F0 (control), F1 (yeasts and lactic acid bacteria), F2 (lactic acid bacteria only), F3 (yeasts only), with evaluation of the microbial succession by plate counting, determination of enzymatic activities by classical methods and qualitative evaluation of flavour compounds by gas-chromatography (GC-MS) with headspace sampling. Besides, QS was estimated by quantification of the expression of luxS genes by Reverse Transcriptase Real Time PCR analysis using selected primers. The results demonstrated that microbial successions were displayed in lab conditions, but no statistical difference in terms of microbial enumeration and α-diversity metrics were observed among the experimental and control fermentations. Moreover, enzymatic activities were not correlated to the total microbiota, indicating the seeds’ endogenous hydrolases protagonist enzymes secretion and activity. Regarding luxS genes measuring for the species Lactiplantibacillus plantarum and Limosilactobacillus fermentum, genes were active in fermentation in the start to the end phase and to the beginning to the middle phase of fermentation, respectively. Correlation analysis among luxS expression and volatile metabolites evidenced Lp. plantarum association with detrimental compounds for fermentation quality. This data contributes to our previous research which monitored fermentations to survey enzymatic changes and QS potential along the process and sheds light of QS-related strategies of lactobacilli dominance in cocoa fermentations.

Competing Interest Statement

The authors have declared no competing interest.

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Investigating interspecific quorum sensing influence on cocoa fermentation quality through defined microbial cocktails
O.G.G. Almeida, M. G. Pereira, R. L. Bighetti-Trevisan, E.S. Santos, E. G. De Campos, G.E. Felis, L.H.S. Guimarães, M.L.T.M Polizeli, B. S. De Martinis, E.C.P. De Martinis
bioRxiv 2022.06.14.496151; doi: https://doi.org/10.1101/2022.06.14.496151
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Investigating interspecific quorum sensing influence on cocoa fermentation quality through defined microbial cocktails
O.G.G. Almeida, M. G. Pereira, R. L. Bighetti-Trevisan, E.S. Santos, E. G. De Campos, G.E. Felis, L.H.S. Guimarães, M.L.T.M Polizeli, B. S. De Martinis, E.C.P. De Martinis
bioRxiv 2022.06.14.496151; doi: https://doi.org/10.1101/2022.06.14.496151

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