Development of a rapid spectrophotometric method for detecting bacterial mucinase complex

A rapid spectrophotometric method for detecting the mucinase complex was developed. Bovine submaxillary mucin is cleaved by commercial mucinase between the oligosaccharide chain and the side chain of peptide linkage, thereby liberating the N-acetyl neuraminic acid (NANA). The release of NANA resulted in an increase of absorbance at 280 nm. The susceptibility to NANA by the new method was found to be at least 10-fold more sensitive than the thiobarbituric acid method. Moreover, the quantification of NANA released from mucin by commercial neuraminidase and partially purified Vibrio parahaemolyticus mucinase showed a good linear correlation in proportion to the concentration of the enzyme used. These results demonstrate that the rapid identification of mucin degradation can be determined by a spectrophotometric assay, thereby providing a new, fast, and sensitive method for assaying the bacterial mucinase complex.