Production of fructooligosaccharides by mycelium-bound transfructosylation activity present in Cladosporium cladosporioides and Penicilium sizovae

Different filamentous fungi isolated from molasses and jams (kiwi and fig) were screened for fructooligosaccharides (FOS) producing activity. Two strains, identified as Penicilium sizovae (CK1) and Cladosporium cladosporioides (CF215), were selected on the basis of the FOS yield and kestose/nystose ratio. In both strains the activity was mostly mycelium-bound. Starting from 600 g/L of sucrose, maximum FOS yield was 184 and 339 g/L for P. sizovae and C. cladosporioides, respectively. Interestingly, the highest FOS concentration with C. cladosporioides was reached at 93% sucrose conversion, which indicated a notable transglycosylation to hydrolysis ratio. The main FOS in the reaction mixtures were identified by HPAEC-PAD chromatography. C. cladosporioides synthesized mainly 1-kestose (158 g/L), nystose (97 g/L), 1F-fructosylnystose (19 g/L), 6-kestose (12 g/L), neokestose (10 g/L) and a disaccharide (34 g/L) that after its purification and NMR analysis was identified as blastose [Fru-beta(2-6)-Glc]. P. sizovae was very selective for the formation of 1F-FOS (in particular 1-kestose) with minor contribution of neoFOS and negligible of levan-type FOS.
Projects BIO2010-20508-C04-01, BIO2010-20508-C04-04, BIO2013-48779-C4-1-R and BIO2013-48779-C4-4-R from Spanish Ministry of Economy and Competitiveness supported this research. We thank Lifelong Learning Programme/Erasmus student placement 2012/2013, and Fundacion Ramon Areces for the institutional Grant to the Centro de Biología Molecular Severo Ochoa.