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Article

Pectinases Secretion by Saccharomyces cerevisiae: Optimization in Solid-State Fermentation and Identification by a Shotgun Proteomics Approach

by
Matheus Mikio Takeyama
1,
Márcia Corrêa de Carvalho
1,
Helena Sacco Carvalho
2,
Cristiane Rodrigues Silva
1,
Ana Paula Trovatti Uetanabaro
3,
Andrea Miura da Costa
3,
Joseph A. Medeiros Evaristo
4,
Fábio César Sousa Nogueira
4,5,
Ana Elizabeth Cavalcante Fai
1,6 and
Maria Gabriela Bello Koblitz
1,*
1
Food and Nutrition Graduate Program (PPGAN), Federal University of the State of Rio de Janeiro (UNIRIO), Rio de Janeiro 22290-240, RJ, Brazil
2
Nutrition School, Federal University of the State of Rio de Janeiro (UNIRIO), Rio de Janeiro 22290-240, RJ, Brazil
3
Department of Biological Sciences, Santa Cruz State University (UESC), Ilhéus 45662-900, BA, Brazil
4
Laboratory of Proteomics/LADETEC, Institute of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro 21941-598, RJ, Brazil
5
Proteomics Unit, Institute of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro 21941-909, RJ, Brazil
6
Department of Basic and Experimental Nutrition, Rio de Janeiro State University (UERJ), Rio de Janeiro 20550-013, RJ, Brazil
*
Author to whom correspondence should be addressed.
Molecules 2022, 27(15), 4981; https://doi.org/10.3390/molecules27154981
Submission received: 20 May 2022 / Revised: 24 June 2022 / Accepted: 25 June 2022 / Published: 5 August 2022
(This article belongs to the Special Issue Food Chemistry: Food Quality and New Analytical Approaches)
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Abstract

A sequential design strategy was applied to optimize the secretion of pectinases by a Saccharomyces cerevisiae strain, from Brazilian sugarcane liquor vat, on passion fruit residue flour (PFRF), through solid-state fermentation (SSF). A factorial design was performed to determine the influence variables and two rotational central composite designs were executed. The validated experimental result was of 7.1 U mL−1 using 50% PFRF (w/w), pH 5, 30 °C for 24 h, under static SSF. Polygalacturonase, pectin methyl esterase, pectin–lyase and pectate–lyase activities were 3.5; 0.08; 3.1 and 0.8 U mL−1, respectively. Shotgun proteomics analysis of the crude extract enabled the identification of two pectin–lyases, one pectate–lyase and a glucosidase. The crude enzymatic extract maintained at least 80% of its original activity at pH values and temperatures ranging from 2 to 8 and 30 to 80 °C, respectively, over 60 min incubation. Results revealed that PFRF might be a cost-effective and eco-friendly substrate to produce pectinases. Statistical optimization led to fermentation conditions wherein pectin active proteins predominated. To the extent of our knowledge, this is the first study reporting the synthesis of pectate lyase by S. cerevisiae.
Keywords: response surface methodology; solid-state fermentation; carbohydrate active enzymes; pectinolytic enzymes; nano LC-MS/MS analysis response surface methodology; solid-state fermentation; carbohydrate active enzymes; pectinolytic enzymes; nano LC-MS/MS analysis

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MDPI and ACS Style

Takeyama, M.M.; de Carvalho, M.C.; Carvalho, H.S.; Silva, C.R.; Uetanabaro, A.P.T.; da Costa, A.M.; Evaristo, J.A.M.; Nogueira, F.C.S.; Fai, A.E.C.; Koblitz, M.G.B. Pectinases Secretion by Saccharomyces cerevisiae: Optimization in Solid-State Fermentation and Identification by a Shotgun Proteomics Approach. Molecules 2022, 27, 4981. https://doi.org/10.3390/molecules27154981

AMA Style

Takeyama MM, de Carvalho MC, Carvalho HS, Silva CR, Uetanabaro APT, da Costa AM, Evaristo JAM, Nogueira FCS, Fai AEC, Koblitz MGB. Pectinases Secretion by Saccharomyces cerevisiae: Optimization in Solid-State Fermentation and Identification by a Shotgun Proteomics Approach. Molecules. 2022; 27(15):4981. https://doi.org/10.3390/molecules27154981

Chicago/Turabian Style

Takeyama, Matheus Mikio, Márcia Corrêa de Carvalho, Helena Sacco Carvalho, Cristiane Rodrigues Silva, Ana Paula Trovatti Uetanabaro, Andrea Miura da Costa, Joseph A. Medeiros Evaristo, Fábio César Sousa Nogueira, Ana Elizabeth Cavalcante Fai, and Maria Gabriela Bello Koblitz. 2022. "Pectinases Secretion by Saccharomyces cerevisiae: Optimization in Solid-State Fermentation and Identification by a Shotgun Proteomics Approach" Molecules 27, no. 15: 4981. https://doi.org/10.3390/molecules27154981

APA Style

Takeyama, M. M., de Carvalho, M. C., Carvalho, H. S., Silva, C. R., Uetanabaro, A. P. T., da Costa, A. M., Evaristo, J. A. M., Nogueira, F. C. S., Fai, A. E. C., & Koblitz, M. G. B. (2022). Pectinases Secretion by Saccharomyces cerevisiae: Optimization in Solid-State Fermentation and Identification by a Shotgun Proteomics Approach. Molecules, 27(15), 4981. https://doi.org/10.3390/molecules27154981

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