Medium engineering of phenylethanoid transfructosylation catalysed by yeast β-fructofuranosidase.

Tyrosol and hydroxytyrosol, by-products of olive oil production, are valuable substrates for enzymatic transglycosylation that can provide products with pharmaceutical potential. Phenylethanoid fructosides are produced from sucrose and phenylethanoids by the catalytic action of β-fructofuranosidases. This work dealt with the potential of the most abundant β-fructofuranosidase, baker's yeast invertase, for this bioconversion.

Design of immobilized biocatalyst and optimal conditions for tyrosol β-galactoside production.

Tyrosol β-galactoside (TG) is a phenylethanoid glycoside with proven neuroprotective properties. This work deals with its biocatalytic production from tyrosol and lactose using Aspergillus oryzae β-galactosidase in immobilized form. Six commercial carriers were examined to find the optimal biocatalyst.

Screening of Commercial Enzymes for Transfructosylation of Tyrosol: Effect of Process Conditions and Reaction Network.

Enzymatic fructosylation of organic acceptors other than saccharides brings new possibilities to synthesize molecules that do not exist in nature. The introduction of fructosyl moiety may lead to glycosides possessing enhanced physicochemical and bioactive properties which could be useful in the pharmaceutical and cosmetic industry. In this work, the regioselective synthesis of tyrosol β-d-fructofuranoside (TF) catalyzed by β-fructofuranosidase is investigated.

Chromatographic purification of recombinant human erythropoietin.

Recombinant human erythropoietin is a valuable therapeutic protein used in the treatment of several serious diseases. It exists in different isoforms and is produced by genetically modified mammalian cells such as Chinese hamster ovary or human embryonic kidney cells. As for other biopharmaceutical drugs, a key factor for its successful industrial production is to achieve a high degree of purity and to decrease the content of critical impurities to trace amounts.

A Method of Early Phase Selection of Carrier for Aspergillus Oryzae β-Galactosidase Immobilization for Galactooligosaccharides Production.

Early phase development of industrial immobilized biocatalysts has to address the selection of the best candidates from dozens of available carriers and binding methods. This work presents a simple selection method for the immobilization of industrial-grade Aspergillus oryzae β-galactosidase suitable for the production of galactooligosaccharides. Immobilization efficiency and yield of a variety of immobilized biocatalysts are evaluated using simple activity measurements and mathematical modeling of intraparticle kinetics and mass transfer.

Application of a micromembrane chromatography module to the examination of protein adsorption equilibrium.

A micromembrane chromatography module based on a 96-well plate design and enabling fast and simple separation of small amounts of proteins was used for the determination of binding capacities of lysozyme, bovine serum albumin, ovalbumin, bovine γ-globulin, and human immunoglobulin G on a hydrophobic membrane Sartobind® Phenyl. Dependence of the binding capacity of the proteins on the ammonium sulfate concentration was examined in the salt concentration range of 0.5-2.

Characterization of pore structure of chromatographic adsorbents employed in separation of monoclonal antibodies using size-exclusion techniques.

Structural properties of commercial chromatographic adsorbents designated for separation of monoclonal antibodies were investigated using size-exclusion techniques. A batch technique provided the specific pore volumes distributed among small, medium and large pores. Inverse size-exclusion chromatography yielded the partition coefficients of dextran solute probes in medium pores.

Chromatographic separation and kinetic properties of fructosyltransferase from Aureobasidium pullulans.

Efficient chromatographic separation of fructosyltransferase from Aureobasidium pullulans was achieved on a preparative scale using a weak anion-exchanger Sepabeads FP-DA. The recovery yield was about 70% and the purification factor reached a value of 28. The molecular weight of the enzyme determined by size-exclusion chromatography was 570,000.

Adsorption equilibrium of fructosyltransferase on a weak anion-exchange resin.

The adsorption equilibrium of a glycoprotein, fructosyltransferase from Aureobasidium pullulans, on an anion-exchange resin, Sepabeads FP-DA activated with 0.1M NaOH, was investigated. The adsorption isotherms were determined at 20 degrees C in a phosphate-citrate buffer with pH 6.