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TitleRecombinant Saccharomyces cerevisiae as a microbial biocatalyst for the one-step production of prebiotic fructooligosaccharides
Author(s)Amorim, Cláudia
Braga, Adelaide
Rodrigues, Joana Lúcia Lima Correia
Cardoso, Beatriz A. Batista
Rainha, João
Gudiña, Eduardo José
Silvério, Sara Isabel Cruz
Rodrigues, L. R.
Issue date26-Nov-2020
CitationAmorim, Cláudia; Braga, Adelaide; Rodrigues, Joana L.; Cardoso, B.; , J. Rainha; Gudiña, Eduardo J.; Silvério, Sara C.; Rodrigues, Lígia R., Recombinant Saccharomyces cerevisiae as a microbial biocatalyst for the one-step production of prebiotic fructooligosaccharides. Biocatalysis Open Day. online, Nov 26, 2020.
Abstract(s)Fructooligosaccharides (FOS) are widely consumed prebiotics with proven beneficial effects on both human and animal health. As a result, alternative production processes with high-efficiency have been an increasing focus of interest by both academy and industry. In this work, a in vivo bioprocess approach was successfully developed for one-step production of FOS from sucrose fermentation by recombinant yeast. Saccharomyces cerevisiae YIL162W lacking the gene responsible for sucrose hydrolysis (suc2) was transformed to express the -fructofuranosidase (Ffase) INV gene from Schwanniomyces occidentalis (clone L196), and its mutated version containing a serine instead of a leucine at position 196 (clone S196), under the inducible GAL1 promotor. Clone S196 presented a 2.75-fold higher sucrolytic activity (22±3 U.mL-1), while clone L196 presented a higher efficiency towards FOS production, producing mainly 6-kestose (76±3 g.L-1) and 1-kestose (1.6±0.6 g.L- 1) after 24 h of fermentation at 30 °C and 200 rpm, in a medium containing 300 g/L of sucrose. Attending the potential of process simplification and cost-reduction, the Ffase INV gene was then expressed under the glyceraldehyde-3-phosphate dehydrogenase (GPD) constitutive promoter (clone GPD L196), resulting in a maximum FOS production of 61±4 g.L-1 ( 56±3 g.L-1 of 6-kestose and 5±31 of fructosylnystose) after 48 h of fermentation using 300 g/L of sucrose. Interestingly, the total amount of undesired glucose and fructose present in the media whenever the maximal FOS production was achieved, was 9 times lower with the GDP promoter (5.5±0.9 g.L-1). The present work demonstrates the high potential of this bioprocess approach for industrial production of prebiotic FOS in a single step. Nevertheless, there is still room for yield improvement in future work, namely through bioprocess optimization.
AccessOpen access
Appears in Collections:CEB - Painéis em Conferências / Posters in Conferences

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