Interfacial Biocatalytic Performance of Nanofiber-Supported β-Galactosidase for Production of Galacto-Oligosaccharides

Molecular distribution, structural conformation and catalytic activity at the interface between enzyme and its immobilising support are vital in the enzymatic reactions for producing bioproducts. In this study, a nanobiocatalyst assembly, β-galactosidase immobilized on chemically modified electrospun polystyrene nanofibers (PSNF), was synthesized for converting lactose into galacto-oligosaccharides (GOS). Characterization results using scanning electron microscopy (SEM) and fluorescence analysis of fluorescein isothiocyanat (FITC) labelled β-galactosidase revealed homogenous enzyme immobilization, thin layer structural conformation and biochemical functionalities of the nanobiocatalyst assembly. The β-galactosidase/PSNF assembly displayed enhanced enzyme catalytic performance at a residence time of around 1 min in a disc-stacked column reactor. A GOS yield of 41% and a lactose conversion of 88% was achieved at the initial lactose concentration of 300 g/L at this residence time. This system provided a controllable contact time of products and substrates on the nanofiber surface and could be used for products which are sensitive to the duration of nanobiocatalysis.

Batch and Fed-Batch Ethanol Fermentation of Cheese-Whey Powder with Mixed Cultures of Different Yeasts

Eight yeast strains of Lachancea thermotolerans, Kluyveromyces marxianus, and Kluyveromyces waltii have been tested for their ability to ferment lactose into ethanol in mashes containing 10% (w/v) cheese whey powder (CWP). The K. marxianus NCAIM Y00963 achieved 3.5% (v/v) ethanol concentration at 96–120 h of fermentation. The ethanol production by the selected lactose-positive strains and the well-known ethanologenic Saccharomyces cerevisiae (Levuline Fb) in mixed culture was also investigated at different CWP concentrations and inoculation techniques in batch mode. The mixed culture in an equal ratio (1:1) of cell counts of K. marxianus and S. serevisiae showed an increase in lactose conversion rate. The two yeast strains in a ratio of 3:1 (three-quarters of K. marxianus and a quarter of S. cerevisiae in a total of 4.5 × 1010 cells) resulted in 72.33% efficiency of lactose bioconversion and 7.6% (v/v) ethanol production at 17.5% (w/v) of CWP concentration. In the repeated inoculation process, with the addition of three-quarter part of 3:1 ratio of mixed culture (3.3 × 1010 cells of K. marxianus) into 150 mL CWP mash at initiation and the rest quarter part (1.2 × 1010 cells of S. cerevisiae) at 24 h, 8.86% (v/v) ethanol content with 87.5% efficiency of lactose conversion was reached. Both the ethanol concentration and efficiency of bioconversion were increased to 10.34% (v/v) and 92%, respectively, by combination with fed-batch fermentation technology. Our results can serve a very good basis for the development of industrial technology for the utilization of cheese whey.

Simultaneous enzymatic hydrolysis and lactic fermentation to obtain a yogurt with low lactose content

In a single-stage process with the simultaneous addition of β-galactosidase and lactic culture, the lactose conversion, the processing time, viscosity and syneresis were evaluated. Fermentation was promoted by lactic culture containing two probiotic microorganisms, Bifidobacterium animalis and Lactobacillus acidophilus, associated with the typical microorganisms of yogurt. An enzymatic preparation containing β-galactosidases from Kluyveromyces lactis and Aspergillus niger was used. A central composite design (2³ trials plus three central points) was proposed in order to evaluate the effects of initial lactose concentration, enzyme concentration and the time of addition of the enzyme. The following conditions were established: initial lactose concentration of 91 g L-1, enzyme concentration of 0.5 g L-1 and enzyme addition at the beginning of fermentation. In these conditions, a decrease in processing time (from 4.55 to 3.68 h) and an increase in lactose conversion (from 15.2 to 97.9%) were observed in relation to the fermentation without enzyme addition, and no detrimental changes in some physical properties of yogurt (viscosity, and syneresis).