Development of a rapid enzymatic quantification method for hyaluronic acid based on the gene-mining of hyaluronate lyase.

Hyaluronic acid (HA), a vital polysaccharide naturally present in human tissues, is widely utilized in the food, pharmaceutical and cosmetic industries due to its diverse functionalities and bioactivity. Rapid and accurate quantification of HA is essential for the quality control of its products. Enzymatic quantification methods, known for their simplicity and high specificity, were employed for polysaccharide measurement. However, the efficient acquisition of enzymes with both high stability and activity posed significant challenges, hindering further advancement in such methods. In this study, we established a novel enzyme screening approach that identified Hly16A, a highly stable and active hyaluronate lyase from the PL16 family. A rapid enzymatic quantification method for HA was established based on Hly16A, demonstrating ideal linearity in the 40-600 μg/mL range. The method enabled HA quantification within 30 min, with no interference from polysaccharides such as chondroitin sulfate and heparin. This study not only provided a rapid and specific method for HA detection, supporting quality control in HA products, but also offered a promising strategy for enzymatic quantification of other polysaccharides, enabled by a gene-mining approach.