title: A simple, rapid and sensitive plate assay for detection of microbial hyaluronidase activity.
abstract: Hyaluronidase (hyase) is a glycosidase enzyme that predominantly degrades hyaluronic acid (HA) having important applications in many biotechnological processes and therapeutics. Several assay methods have been proposed to screen hyase producing microorganisms; however, they rely on unique reagents and sophisticated instruments, which are expensive and could be unavailable in general laboratories. In the present studies, a rapid, simple, sensitive, highly reproducible, and cost-effective qualitative plate assay has been developed for the screening of hyase producing microorganisms. The routinely used plate assay method of Richman and Baer requires a special chemical cetylpyridinium chloride and long incubation period of 20 h; but still, the zones of clearance are not very clear and distinct. While, the present method requires an incubation period of only 1 h and the distinct zones of clearance appear with Gram's iodine within 1 min of time. This method does not require any special medium, unlike previously reported methods. Moreover, use of commonly available Gram's iodine makes this method suitable for many researchers. The results of the assay method were validated by TLC, zymographic analysis and determining the growth of isolates in minimal medium containing HA as a sole carbon source.
authors.affiliation: Department of Microbiology and Biotechnology, R.C. Patel Arts, Commerce and Science College, Shirpur, India.
authors.forename: Sandip
authors.initials: S
authors.lastname: Patil
authors.affiliation: Department of Microbiology, School of Life Sciences, North Maharashtra University, Jalgaon, India.
authors.forename: Bhushan
authors.initials: B
authors.lastname: Chaudhari
mesh_terms: D003470:Culture Media / Q000737:chemistry; D057075:Enzyme Assays / Q000191:economics / Q000379:methods*; D006801:Humans; D006820:Hyaluronic Acid / Q000302:isolation & purification / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000302:isolation & purification* / Q000378:metabolism; D007455:Iodine; D012680:Sensitivity and Specificity; D012685:Sepharose; D018502:Streptococcus equi / Q000737:chemistry / Q000201:enzymology* / Q000254:growth & development; D034361:Streptococcus mitis / Q000201:enzymology
chemical_list: D003470:Culture Media; D006820:Hyaluronic Acid; D012685:Sepharose; D007455:Iodine; D006821:Hyaluronoglucosaminidase
keywords: Gram's iodine; hyaluronidase; plate assay