Quercetin is a flavonoid largely employed as a phytochemical remedy and a food or dietary supplement. We present here a novel biocatalytic methodology for the preparation of quercetin from plant-derived rutin, with both substrate and product being in mostly an undissolved state during biotransformation. This “solid-state” enzymatic conversion uses a crude enzyme preparation of recombinant rutinosidase from Aspergillus niger yielding quercetin, which precipitates from virtually insoluble rutin. The process is easily scalable and exhibits an extremely high space-time yield. The procedure has been shown to be robust and was successfully tested with rutin concentrations of up to 300 g/L (ca 0.5 M) at various scales. Using this procedure, pure quercetin is easily obtained by mere filtration of the reaction mixture, followed by washing and drying of the filter cake. Neither co-solvents nor toxic chemicals are used, thus the process can be considered environmentally friendly and the product of “bio-quality.” Moreover, rare disaccharide rutinose is obtained from the filtrate at a preparatory scale as a valuable side product. These results demonstrate for the first time the efficiency of the “Solid-State-Catalysis” concept, which is applicable virtually for any biotransformation involving substrates and products of low water solubility.

The present study describes the production of the value-added chemical 1,3-propanediol (1,3-PD) from crude glycerol, a waste by-product formed during biodiesel production. The efficiency, robustness, and stability of the process were improved by immobilization of the anaerobic bacterium Clostridium butyricum into a polyvinyl alcohol (PVA) hydrogel. The highest average productivity, 6.8 ± 0.2 g/(L·h), was achieved in 10 consecutive, repeated batch fermentations, with an initial concentration of pure glycerol 45.5 ± 0.7 g/L, after 2.5 h. The highest final concentration and yield of 1,3-PD, 28.3 ± 0.6 g/L, and 0.42 ± 0.01 g/g, respectively, were achieved in eleven repeated batch fermentations, after increasing the initial pure glycerol concentration to 70.4 ± 1.9 g/L. Two different types of crude glycerol, produced from used cooking oil (UCO) and rapeseed oil (RO), were tested in repeated batch fermentations, with an average productivity achieved of 2.3 ± 0.1 and 3.5 ± 0.3 g/(L·h), respectively. The highest final concentration and yield of 1,3-PD, 12.6 ± 0.9 g/L, and 0.35 ± 0.02 g/g, respectively, were observed in fifteen repeated batch fermentations with RO crude glycerol. An excellent stability of the immobilized anaerobic bacteria and increase of productivity in fermentation of crude glycerol was demonstrated.

Arylmalonate decarboxylase (AMD) is a monomeric enzyme of only 26 kDa. A recombinant AMDase from Bordetella bronchiseptica was expressed in Escherichia coli and the enzyme was immobilized using different techniques: entrapment in polyvinyl alcohol (PVA) gel (LentiKats^®), covalent binding onto magnetic microparticles (MMP, PERLOZA s.r.o., Lovosice, Czech Republic) and double-immobilization (MMP-LentiKats^®) using the previous two methods. The double-immobilized AMDase was stable in 8 repeated biocatalytic reactions. This combined immobilization technique has the potential to be applied to different small proteins.