Significance of Debranching Enzyme (DBE) Gene Modification

Plants contain an α(1→6) glucan hydrolase, starch debranching enzyme (DBE), which is mainly involved in the synthesis of amylopectin and acts as a modifier of the structure of starch. Depending on the mode of action on the substrate, DBE is categorized as pullulanase (also known as limit dextrinase or R-enzyme) and isoamylase. Due to its ability to hydrolyze the branching bonds in amylopectin, DBE is also used in large quantities in the starch industry to prepare resistant starch, cyclodextrins, and so on.

Since the co-catalysis of DBE with other starch-acting hydrolases can effectively improve the raw material utilization and productivity in starch processing steps (e.g., saccharification and modification), it has attracted extensive research interest. Natural DBE has low catalytic efficiency and poor thermal stability. In order to improve the performance of DBE, it can be genetically modified for targeted modification of its catalytic efficiency and thermal stability to expand its application range.

Innovative Services

CD BioGlyco has a lot of experience in gene modification, and we modify the DBE gene in several flowing directions:

• DBE gene-based expression upregulation service
  Over-expression vector is constructed, and transgenic lines are constructed by the Agrobacterium method. After cultured for a certain time to analyze DBE specific expression, DBE activity is detected, and starch content synthesized in transgenic plants is compared, starch granules are observed.

• DBE gene-based expression suppression service
  Using RNAi technology, we construct RNAi vectors to inhibit the expression of the DBE gene and introduce the vectors into plant healing tissues. Then we will test the activity of DBE after cultivation for a certain period, analyze the expression amount of DBE, and detect the differences in starch content, branched starch chain length distribution, etc., between natural and transgenic plants.

• Targeted mutations service
  Based on the sequence comparison and protein simulation analysis, the DBE coding gene is targeted mutated, and DBE mutant lines are constructed. After a fixed period of cultivation, DBE activity is measured. The amino acid sequences of the natural and mutant DBE will be analyzed and compared. The three-dimensional structure of mutant DBE will be simulated. We also analyze the specific expression of the DBE gene and detect the changes in the structure of amylopectin, etc.

Fig.1 Experimental Procedure. (CD BioGlyco)

Applications

• DBE plays an important role in plant metabolism. Researchers used gene editing techniques to explore the role of DBE in sugar anabolism.
• Gene editing techniques can influence starch synthesis in crops. It enables researchers to explore crops that produce excellent-quality grain.
• DBE gene editing technology is used to study the role of DBE in seed development, and plant growth.

Advantages

• Our method of screening positive strains is very effective. The shape is stable and does not recover.
• Our gene editing technology has been optimized. High success rate of gene editing and low off-target rate.
• Multiple gene editing protocols are available. Our customized gene editing service meets different research needs.

CD BioGlyco focuses on optimizing gene-editing services to modify starch. We construct different vectors according to customers' needs and provide various products and analytical techniques to help accelerate scientific research. Besides, we also provide Physical, Chemical, and Enzymatic Modification services. If you have any needs, please feel free to contact us.

• ADP-Glucose Pyrophosphorylase (AGPase) Modification
• Starch Synthase Modification
• Starch-Branching Enzyme Modification