Applications of Enzymatic Degradation/Hydrolysis

Enzymatic degradation/hydrolysis or its combination with other processes is an effective way to convert lignocellulosic materials into biofuels or other value-added products. The use of enzymatic degradation/hydrolysis typically involves pretreatment followed by microbial or chemical conversion as a process to release monosaccharides from cellulose and hemicellulose. Importantly, enzymatic degradation/hydrolysis can generate oligosaccharides in addition to sugar monomers. For example, cello-oligosaccharide (COS) in which cellobiose is the main oligosaccharide unit can be obtained by enzymatic degradation/hydrolysis of cellulose. Although COS has not received as much attention as potential prebiotics as hemicellulose-derived oligosaccharides such as arabinooligosaccharides, some studies have shown that it can provide multiple positive functions for host health, including affecting the intestinal mucosal structure, absorption function, barrier integrity, and gut microflora.

Fig.1 Substrate specificity of cell wall degrading enzymes toward the main cell wall polysaccharides. (Giovannoni, et al., 2020)

Enzymatic Degradation/Hydrolysis for Oligosaccharide Manufacture at CD BioGlyco

CD BioGlyco provides satisfactory 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. Based on these advanced solutions and processes, we provide advanced oligosaccharide manufacturing services, including Galactooligosaccharide (GOS), lacto-sucrose, Xylo-oligosaccharide (XOS), COS, mannano-oligosaccharide (MOS), and arabino-XOS (AXOS). Our enzymatic hydrolysis/degradation service is divided into the following steps.

• Raw material treatment

Raw materials are selected and processed according to one or several steps of hammer milling, alkali dissolution, acid hydrolysis, water bath, filtration, washing, drying, and so on.

• Enzyme expression and purification

According to the experimental requirements and test conditions, we will obtain the corresponding enzymes for the experiment.

• Determination of enzyme protein concentration and activity

Protein concentration is determined by the Bradford method with bovine serum albumin (BSA) as the calibration standard. According to the recommendation of IUPAC, we select a certain substance as the substrate to measure enzyme activity.

• Enzymatic hydrolysis

Using the selected enzyme alone in a suitable test tube to hydrolyze the substrate. If necessary, we will choose liquid hot water pretreatment to improve the efficiency of enzymatic hydrolysis. At the optimal temperature and pH, the enzyme and substrate are mixed and shaken for a certain period in a shaker incubator. After the reaction, configure the buffer to adjust the pH of the final solution in each tube.

• Quantification of oligosaccharides

The content of oligosaccharides in the supernatant of the enzymatic hydrolysis mixture is determined by chromatography. Standards are used to construct a calibration curve for the quantitative determination of oligosaccharides in samples.

Fig.2 Oligosaccharides produced by enzymatic degradation/hydrolysis. (CD BioGlyco)

Applications

• Advances in enzymatic hydrolysis/degradation facilitate the development of enzyme reaction engineering practices, custom-made enzyme preparations, controlled enzymatic hydrolysis, and protein engineering approaches.
• Enzymatic hydrolysis/degradation is an important method for the production of oligosaccharides.
• The development of enzymatic hydrolysis/degradation technology is beneficial to the production of bioenergy.

Advantages

• Enzymatic hydrolysis/degradation produces fewer by-products in the production of oligosaccharides.
• Enzymatic hydrolysis/degradation requires mild operating conditions, which increases the safety of the process.
• Enzymatic hydrolysis/degradation allows for enzyme recovery, which reduces costs.

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. Don't hesitate to contact us if you're interested in learning more.

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• Enzymatic Epimerization for Oligosaccharide Manufacture