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[Lipase activity of Oospora lactis protoplasts]
Mikrobiologiia
|May 1, 1980
Summary
Researchers developed a method to create osmotically sensitive fungal protoplasts from Oospora lactis using lytic enzymes. This process efficiently releases periplasmic lipase without altering its molecular characteristics.
Area of Science:
- Mycology
- Enzymology
- Biochemistry
Context:
- Fungal cell wall lysis is crucial for protoplast isolation.
- Oospora lactis is a fungus with potential biotechnological applications.
- Enzymatic degradation offers a targeted approach to cell wall disruption.
Purpose:
- To describe a reproducible method for preparing osmotically sensitive protoplasts from Oospora lactis.
- To investigate the enzymatic lysis of Oospora lactis cell walls.
- To characterize the release and properties of periplasmic lipase during protoplast formation.
Summary:
- Oospora lactis cells were effectively lysed to form protoplasts using a lytic enzyme from actinomycetes at specific concentrations, temperature, and pH.
- Optimal stabilization of protoplasts was achieved with 1 M NaCl or a 1:1 mixture of 1 M NaCl and 1 M mannitol.
- Pretreatment with Tris, EDTA, and cysteine was necessary for cell wall lysis induced by Helix pomatia enzyme.
- Protoplast formation led to the release of 90% of intracellular lipase activity from the periplasmic space.
- Exocellular and periplasmic lipase exhibited identical electrophoretic mobility, pH optimum, and substrate specificity, indicating no molecular weight change upon release.
Impact:
- Establishes a reliable protocol for Oospora lactis protoplast generation, facilitating further genetic and biochemical studies.
- Demonstrates the utility of specific lytic enzymes and stabilizing agents in fungal protoplast isolation.
- Provides insights into the localization and release mechanism of lipase in Oospora lactis.
- The characterization of released lipase supports its potential use in biotechnological applications.