Preparatory production of quercetin-3-β-d-glucopyranoside using alkali-tolerant thermostable α-l-rhamnosidase from Aspergillus terreus

Extensive screening for a robust producer of α-l-rhamnosidase activity from well-defined strains of filamentous fungi, including multifactorial optimization (inducers, cultivation conditions) was accomplished. Enzyme production of the optimal producer Aspergillus terreus (non-toxigenic) was scaled up to 50 L. α-l-Rhamnosidase, which was fully characterized, proved to be thermo- and alkali-tolerant, thus enabling effective operation at 70 °C and pH 8.0. These conditions allow for a very high substrate (rutin) load up to 100–300 g/L, thus enabling very high volumetric productivity of the reaction product quercetin-3-β-d-glucopyranoside (isoquercitrin). Here, a novel concept of “immobilised substrate” is used. Isoquercitrin is a highly effective and biocompatible antioxidant with strong anti-inflammatory activities. Rutin biotransformation was optimized and scaled up to ca 10 kg production and thus the robustness of the large-scale production was demonstrated. Isoquercitrin can be produced to a very high purity (98%) in multikilogram amounts, without any quercetin and directly applicable in nutraceuticals.

Figure optionsDownload as PowerPoint slideHighlights
► A new robust thermo- and alkalitolerant extracellular α-l-rhamnosidase from Aspergillus terreus was found.
► Rutin was converted into quercetin-3-β-d-glucopyranoside (isoquercitrin) by the biotransformation with α-l-rhamnosidase.
► Biotransformation procedure was scaled up to 30 L and up to volumetric productivity of over 150 g/L.
► A new concept of the “immobilized substrate”, e.g. bioconversion in the solid state substrate was developed.
► The procedure is waste-free; the liberated rhamnose was recycled for the enzyme production in the fermentation.