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Biodegradation of Griseofulvin by Bacillus subtilis isolated from expired pharmaceuticals raw materials
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Biodegradation of Griseofulvin by Bacillus subtilis isolated from expired pharmaceuticals raw materials


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Aim of the Presentation …

Aim of the Presentation
1. Study the biodegradation process of pharmaceutical raw materials.
2. Purification of the biodegradation enzymes.
3. Identification of the biodegradation products.

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  • 1. By Dr. Yahia Ahmed Maher Assistant professor of Microbiology Biodegradationof Griseofulvinby Bacillussubtilis isolatedfromexpiredpharmaceuticalsraw materials Qura University-Faculty of Dentistry Umm Al  
  • 2. Aim of the Work 2 Purification of the biodegradation enzymes. 3 Identification of the biodegradation products. Study the biodegradation process of pharmaceutical raw materials.1
  • 3. • The pharmaceutical industry products are present in everyday life, for a long time the production of chemicals and pharmaceuticals as well as their usage and application caused heavy pollution of the environment and serious health effects (Sheldon, 2007). Introduction
  • 4. • Biodegradation is a very important process in the transformation of organic pollutants as pharmaceuticals by bacterial growth as energy source during metabolically processes that helps to maintain ecosystem and human health (Yu and Wu, 2012). Introduction
  • 5. • Some microorganisms may cause drug contaminant and unable to replicate but could remain viable and cause lowering of the quantity of the therapeutic agent in the dosage form and decrease in its bioavailability or released a toxic product into environment such as phenol and acids (United States Pharmacopoeia 2004). Introduction
  • 6. • Griseofulvin is antifungal produced by growth of certain strains of Penicillium griseofulvum and used against dermatophytic infection, young actively metabolized cells and inactive against Actinomyces, Nocardia, and other bacteria (United States Pharmacopoeia 2004). Introduction
  • 7. Structure formula of Griseofulvin. Introduction
  • 8. 1   • Sample collection. 2   • Culture medium and growth conditions. 3   • Isolation, purification and identification. 4   • Studying the optimum condition for enzyme production. 5   • Detection of degrading Griseofulvin enzyme. 1- Study the biodegradation processes
  • 9. 2- Griseofulvin identification method A- Physical test B- Chemical reaction C- Spectroscopical analysis a- UV spectroscopy detection c- IR detectionb- TLC detection d- HPLC detection
  • 10. 1   • Filtration the broth and precipitatedby using ammonium sulfate. 2   • Dialysis. 3   • Purifiedby using DEAE-Cellulose, sephadex G-200 column chromatography and SDS-polyacrylamide gel electrophoresis. 4   • Identificationof the Griseofulvinbiodegradationproducts by using GC-Mass Spectrometry. 3- Extraction and purificationof Griseofulvin biodegradation
  • 11. Sample collection • Eleven bacterial isolates from six expired pharmaceutical raw materials on Tryptic Soy Agar (TSA) and Nutrient Agar (NA) medium from total different seventy-eight samples of expired pharmaceutical raw materials which collected from the storage room of Memphis Company for pharmaceutical and chemical industry.
  • 12. Contaminated and non-contaminated expired pharmaceutical raw materials
  • 13. Microphotograph of Bacillus subtilis
  • 14. Detection of degrading Griseofulvin enzyme
  • 15. Griseofulvin cup plate clearing zone (CZ) technique of Griseofulvin degrading enzyme
  • 16. 2- Griseofulvin identification method • The physical, chemical and UV spectroscopy don’t detect any degradation for Griseofulvin while it is detected by TLC, IR and HPLC.
  • 17. TLC recording visible estimation of standard Griseofulvin ST T
  • 18. IR spectrum analysis of standard and biodegraded Griseofulvin
  • 19. HPLC analyses of standard and biodegraded Griseofulvin 92.85%
  • 20. Standard marker and purified Griseofulvin degrading enzyme A, standard marker lane. B, Griseofulvin degrading enzyme lane (5.3439 KDa).
  • 21. Relativeabundance Retention time (Min) GC-MS detection of standard Griseofulvin Griseofulvin
  • 22. Relativeabundance Retention time (Min) GC-MS detection of biodegradation Griseofulvin Diethyl phtalatate CO2C2H5 CO2C2H5 Griseofulvin
  • 23. Conclusion The biodegradation processes require present adapted isolate to synthesize appropriate enzyme. The biodegradation products were extracted and identified by GC-Mass Spectrometry. Remediation of the pharmaceutical raw materials wastes by using biodegradation technique.