Lactulose and Its Manufacture

Lactulose is an indigestible disaccharide that is used medically in the treatment of portal-systemic encephalopathy, chronic constipation, and hyperammonemia. In the food industry, the bifidus factor lactulose can be used as an ingredient with greater sweetness and solubility than lactose.

Cellobiose 2-epimerase has the ability to isomerize the glucose moiety in cellobiose or glucose, so this enzyme has a novel function of converting lactose to lactulose in industrial applications of enzymes. There are some advantages to using thermostable enzymes due to increased substrate solubility and enzyme reaction speed at elevated temperatures. Thus, a thermostable cellobiose 2-epimerase can be used to efficiently convert lactose to lactulose as a single substrate.

Fig.1 Enzymatic synthesis of non-digestible oligosaccharides. (Vera, et al., 2021)

Enzymatic Isomerization for Oligosaccharide Manufacture at CD BioGlyco

CD BioGlyco provides complete One-Stop Solutions for Carbohydrate Manufacture, and our experts customize experimental schemes according to clients' scientific research projects. We offer a first-class and cost-effective Enzymatic Process for Oligosaccharide Manufacture to help clients accelerate their projects. We offer high-quality enzymatic isomerization for oligosaccharide manufacture to maximize the production of lactulose. In this process, lactose is the single substrate and cellobiose 2-epimerase is used for oligosaccharide production. Our enzymatic isomerization process is divided into the following steps.

• Enzyme acquisition

Select appropriate bacterial strains, Escherichia coli, and plasmids for the source of cellobiose 2-epimerase genomic DNA, host cells, and expression vectors, respectively.

• Enzyme gene cloning and purification

Gene cloning of cellobiose 2-epimerase is performed. Recombinant cells are harvested from the culture medium by centrifugation and resuspended in phosphate-buffered saline. After a series of purification operations, a fast protein liquid chromatography system is used for subsequent purification.

• Enzyme assay

The blind sample is used as a control and incubated for the same incubation time with the heat-inactivated enzyme obtained by boiling the enzyme and then the blind sample is prepared after incubation with lactose and no lactose.

• Production of lactulose by cellobiose 2-epimerase

Determine the enzyme concentration for maximum lactulose production by testing enzyme activity at different enzyme concentrations. The effect of substrate concentration on lactulose production is tested by testing different lactulose concentrations. At the same time, the time for cellobiose 2-epimerase to produce lactulose is recorded.

• Analysis

The enzymatic reaction is terminated by adding a certain concentration of HCl to the reaction solution. Subsequently, the samples are diluted to adjust the sugar concentration to a certain concentration. The lactulose concentration in the resulting solution is determined by a high-performance liquid chromatography system.

Fig.2 Enzymatic isomerization process. (CD BioGlyco)

Applications

• Enzymatic isomerization is an important technology for the production of lactulose.
• Enzyme isomerization has the characteristics of clean production and easy purification, and it is an alternative method to chemical synthesis.
• The cellobiose 2-epimerase used in the enzymatic isomerization is thermostable and catalyzes the reaction at high temperatures.

Advantages

• Enzymatic isomerization is an efficient way to synthesize lactulose from the single substrate lactose.
• Due to the cumbersome chemical synthesis process, enzymatic synthesis is an attractive technical choice.
• Protein engineering technology allows to obtain active and stable enzymes.

CD BioGlyco has first-class technology and well-trained technicians. Our dedicated team of experts works tirelessly to provide Oligosaccharide Manufacture Service that meets clients' needs and requirements. For further details, please don't hesitate to contact us.

• Enzymatic Degradation/Hydrolysis for Oligosaccharide Manufacture
• Enzymatic Synthesis for Oligosaccharide Manufacture
• Enzymatic Transglycosylation for Oligosaccharide Manufacture
• Enzymatic Epimerization for Oligosaccharide Manufacture