Design of combined crosslinked enzyme aggregates (combi-CLEAs) of β-galactosidase and glucose isomerase for the one-pot production of fructose syrup from lactose

A new bi-enzymatic catalyst has been produced by precipitation and crosslinking (combi-CLEAs) of β-galactosidase and glucose isomerase for catalyzing the cascade reactions of lactose conversion into fructose, producing a lactose-fructose syrup (LFS). Glucose isomerase was chemically aminated to increase its reactive surface groups for favour the crosslinking step. The effect of β-galactosidase to glucose isomerase activity ratio and glutaraldehyde to protein mass ratio in combi-CLEAs production was evaluated. The selected combi-catalyst was successfully used in the production of fructose syrup from lactose in a single reaction vessel. The biocatalyst could be used at least in five sequential batches of LFS production, remaining fully stable after a total of 50 h of reaction, obtaining a product of constant quality. A robust bi-enzymatic catalyst was produced that can be repeatedly used in LFS production, an attractive mild sweetener for the dairy food industry.