Chitinase Assay

Quantifying chitinase activity in microbial isolate

1. Inoculate a single isolated colony to isolate appropriate liquid rich media.
2. Incubate for 48 h at 25^o^C with shaking.
3. After incubation adjust culture to OD~600nm~ 0.1
4. Place 4-5 ul drops of adjusted culture in four quadrants of a chitin agar plate, spaced equidistant from each other.
   • NOTE: Unless unavoidable, do not inoculate different isolates onto the same plate.
5. Incubate plates at 28^o^C for 5 days.
6. After 5 days, observe colonies for clearing zones around colonies.
7. Measure diameter of all observed clearing zones.

Citations

• Sasi et al 2020. Identification and Characterization of a Newly Isolated Chitinase-Producing Strain Bacillus licheniformis SSCL-10 for Chitin Degradation
• Hsu and Lockwood 1975. Powdered Chitin Agar as a Selective Medium for Enumeration of Actinomycetes in Water and Soil
• Veliz et al 2017. Chitinase-producing bacteria and their role in biocontrol
• Bhattacharya et al 2008. Bacterial Chitinases: Properties and Potential
• Jog et al 2014. Mechanism of phosphate solubilization and antifungal activity of Streptomyces spp. isolated from wheat roots and rhizosphere and their application in improving plant growth
• Gooday. 1990. Advances in Microbial Ecology: The ecology of chitin degradation