Enzyme Activity Measurement for Unsaturated Rhamnogalacturonyl Hydrolase

Creative Enzymes is a leader of high-quality bio-analytical services. We provide personalized contract research specialized in enzyme activity measurement such as the spectrophotometric activity assay for unsaturated rhamnogalacturonyl hydrolase. Due to the recent discovery and identification of this enzyme, little research has been performed to investigate its kinetic behavior and structure. Properly measuring its activity is even harder without the above-mentioned information. However, Creative Enzymes depends on its own superb expert team and developed robust activity analysis to satisfy customer’s request.

Unsaturated rhamnogalacturonyl hydrolase (EC 3.2.1.172, YesR) was recently identified from Bacillus subtilis and categorized into a new glycoside hydrolase family 105 (GH-105) according to their enzyme properties. YesR acts specifically on unsaturated rhamnogalacturonan (RG) and yields ΔGalA from the substrate. The enzyme is suspected to be conserved in bacteria and fungi. Its 3-dimensional structure remains unclear.

Rhamnogalacturonan (RG) resembles the ramified region of pectin, the most complex polysaccharide that composes the plant cell wall. Plant cell walls play critical roles in maintaining cell shape, transporting nutrients, exporting toxins and metabolites, and defending from microbial pathogens. Plant cell wall-degrading enzymes are crucial for microbial pathogens to invade plant cells. Many plant pathogenic bacteria produce enzymes that are in charge of the degradation of RG-I, the main chain of rhamnogalacturonan. The enterobacterium Erwinia chrysanthemi which is a typical member of plant pathogens, causes soft rot diseases and produces different types of RG-I-related enzymes including unsaturated rhamnogalacturonyl hydrolase. For its unique activity, unsaturated rhamnogalacturonyl hydrolase has great potential in food processing, utilization of biomass, and agricultural applications as catalysts or potential target to treat plant pathogens. Therefore, the prospective market of the enzyme is to grow rapidly. However, the standard activity assay for this enzyme has not been established.

Figure 1: Proposed catalytic reaction mechanism of YteR.
Reference: Itoh T, Ochiai A, Mikami B, et al. Biochemical and biophysical research communications, 2006, 347(4): 1021-1029.

Creative Enzymes is proud of providing the precise enzymatic activity assay for unsaturated rhamnogalacturonyl hydrolase. The enzyme activity is measured by monitoring the decrease in absorbance at 235 nm; this corresponded to the loss of the C=C double bond of the substrate. Our test results are the most trusted by thousands of customers who have been enjoying our services for many years. With a long history of research and development, Creative Enzymes has accumulated extensive experiences and become the first choice in enzyme activity measurement.

Figure 2: The crystal structure of YteR protein from Bacillus subtilis.
PDB: 1NC5