Pectinases secretion by Saccharomyces cerevisiae: optimization in solid state fermentation and identification by shotgun proteomics approach.

CORREA DE CRAVALHO, MÁRCIA; MATHEUS MIKIO TAKEYAMA; HELENA SACCO CARVALHO; CRISTIANE RODRIGUES SILVA; JOSEPH MEDEIROS EVARISTO; ANDREA MIURA DA COSTA; FABIO CESAR SOUSA NOGUEIRA,; TROVATTI UETANABARO, ANA PAULA; MARIA GABRIELA BELLO KOBLITZ

MOLECULES

MOLECULAR DIVERSITY PRESERVATION INTERNATIONAL-MDPI

Lugar: Basel; Año: 2022

1420-3049

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). Methods and Results: 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. Conclusions: 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. Significance and Impact of the Study: to the extent of our knowledge, this is the first study reporting the synthesis of pectate lyase by S. cerevisiae.