High-efficient production of fructo-oligosaccharides from inulin by a two-stage bioprocess using an engineered Yarrowia lipolytica strain

Technological Aspects of the Production of Fructo and Galacto-Oligosaccharides. Enzymatic Synthesis and Hydrolysis

“…In a similar process, but in a two-step way, Han et al (92) achieved similar results by using a recombinant Yarrowia lipolytica strain Enop56, in which an optimized endo-inulinase gene from Aspergillus niger was overexpressed. The hydrolysis in these conditions lead to the formation of FOSs with DP 3–5 as major products and to <5% of mono- and disaccharides (non-prebiotic).…”

Hydrolytic secretome engineering in Yarrowia lipolytica for consolidated bioprocessing on polysaccharide resources: review on starch, cellulose, xylan, and inulin

“…No information on testing the SUC2-overproducing Y. lipolytica strains toward inulin were reported there. This issue was however raised in the following study (Han et al 2017) showing that this particular invertase (SUC2) produced by Y. lipolytica shows only marginal specificity toward inulin and fructooligosaccharides. It was determined that the hydrolytic activity of the SUC2 invertase toward fructooligosaccharides was only 8.8% of that toward sucrose, and there was no detectable hydrolytic activity toward inulin (Han et al 2017).…”

“…The two proteins were overproduced separately and their activities were analyzed independently and in combination, demonstrating high synergy in their action (Liu et al 2016). The same expression host was used in another study, where two copies of A. niger endo-inulinase were cloned and overexpressed under strong growth-phase-dependent promoter (Han et al 2017). However, in this approach, the host system was actually used in situ for inulin decomposition, which was executed by a two-stage approach.…”

“…Notably, in the latter stage, the temperature was raised from initial 28 to 35°C, which is more suited for the enzyme's action. Consequently, the authors developed a highly efficient process of fructooligosaccharide production, using recombinant Y. lipolytica strain overproducing two fungal endo-inulinases, and SU2 invertase (Po1h background), which removed any non-prebiotic saccharides generated during the process (Han et al 2017). The abovementioned previous studies on inulinolytic Y. lipolytica (Zhao et al 2010;Liu et al 2016) were finalized by a work on construction of a strain overproducing both exoand endo-inulinase simultaneously, able to accumulate 48.13% (g/gDCW) from inulin (Shi et al 2018).…”

“…The process models use a renewable polysaccharide-inulinand citric acid as the target molecule. All the presumptions were based on literature data on inulin-based bioprocesses, citric acid synthesis from inulin (Han et al 2017;Singh et al 2018;Rakicka et al 2019), and market search for industrial materials prices (MP). In comparison to the simulated CBP process, the cocktail-based process encompasses additional procedure P1b (Fig.…”