Bacteria chitinase production and activity
Faculty Mentor
Charles Herr
Document Type
Poster
Start Date
10-5-2023 9:00 AM
End Date
10-5-2023 10:45 AM
Location
PUB NCR
Department
Biology
Abstract
Fungal contamination is a detrimental problem in cell cultures. The use of antimycotic is inefficient due to their short stability in culture and possible influence on cell activities. The employment of chitinase enzymes in cell cultures then would be the answer owing to their specificity to chitin. Chitin is a major component in fungal cell wall structural stability, comprising chains of monomers/oligomers. Chitinase and related enzymes are produced by many organisms for various purposes, which can be harvested for laboratory uses. Bacteria species were screened on Lysogeny Broth (LB) Agar supplemented with colloidal chitin and cultured for chitinase production. Their chitinase activities were assayed using modified Schales procedure, reflecting chito-monomers/oligomers concentration through Optical Density (O.D.) readings with a Spectrophotometer. Colloidal chitin used for screening and culturing was also assayed on the same procedure to ensure it is free of contamination. Culture broth was observed to influence the assay despite absence of chito-monomers/oligomers. For future experiments, LB Broth will be dialyzed to extract chitin reducing ends. Bacillus subtilis was the model bacterium but observed to have low chitinase activity in previous experiments. Therefore, a new model bacterium, Serratia marcescens, was screened and chosen for culture and enzyme production. S. marcescens chitinase activity will be assayed with Schales procedure, and O.D. readings will be plotted and compared to purified chitinase from Aspergillus niger. S. marcescens chitinase will also be tested on various substrates to examine chitinase specificity. Our result will enable us to study the potential for laboratory use of chitinase extracted from bacteria cultured in the lab.
Recommended Citation
Nguyen, Nguyen, "Bacteria chitinase production and activity" (2023). 2023 Symposium. 57.
https://dc.ewu.edu/srcw_2023/res_2023/p1_2023/57
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Bacteria chitinase production and activity
PUB NCR
Fungal contamination is a detrimental problem in cell cultures. The use of antimycotic is inefficient due to their short stability in culture and possible influence on cell activities. The employment of chitinase enzymes in cell cultures then would be the answer owing to their specificity to chitin. Chitin is a major component in fungal cell wall structural stability, comprising chains of monomers/oligomers. Chitinase and related enzymes are produced by many organisms for various purposes, which can be harvested for laboratory uses. Bacteria species were screened on Lysogeny Broth (LB) Agar supplemented with colloidal chitin and cultured for chitinase production. Their chitinase activities were assayed using modified Schales procedure, reflecting chito-monomers/oligomers concentration through Optical Density (O.D.) readings with a Spectrophotometer. Colloidal chitin used for screening and culturing was also assayed on the same procedure to ensure it is free of contamination. Culture broth was observed to influence the assay despite absence of chito-monomers/oligomers. For future experiments, LB Broth will be dialyzed to extract chitin reducing ends. Bacillus subtilis was the model bacterium but observed to have low chitinase activity in previous experiments. Therefore, a new model bacterium, Serratia marcescens, was screened and chosen for culture and enzyme production. S. marcescens chitinase activity will be assayed with Schales procedure, and O.D. readings will be plotted and compared to purified chitinase from Aspergillus niger. S. marcescens chitinase will also be tested on various substrates to examine chitinase specificity. Our result will enable us to study the potential for laboratory use of chitinase extracted from bacteria cultured in the lab.
