Rapid method for preparing cell-free extracts of Aspergillus ochraceus

Area of Science:

• Biochemistry
• Mycology
• Enzymology

Background:

• Cell-free extracts are crucial for studying fungal enzyme systems.
• Traditional methods for preparing fungal extracts can be time-consuming and may compromise enzyme activity due to heat or heavy metal exposure.

Purpose of the Study:

• To develop a rapid, efficient, and gentle method for preparing cell-free extracts from Aspergillus ochraceus.
• To ensure the integrity of fungal enzymes for subsequent in vitro studies.

Main Methods:

• Aspergillus ochraceus mycelial mats were rapidly frozen in liquid nitrogen.
• Frozen mycelia were ground into a fine powder using a cold mortar and pestle.
• The powdered mycelia were homogenized in an all-glass mechanical homogenizer at low temperatures.

Main Results:

• The developed method yielded an average of 43.0 mg of protein per gram of mycelium (wet weight).
• The preparation process maintained extract temperatures below 1°C and avoided contact with heavy metals.
• Key enzyme activities, including amylase, acid phosphate dehydrogenase, beta-glucosidase, beta-fructofuranosidase, and trehalase, were successfully measured in the cell-free extracts.

Conclusions:

• The new method is a fast and efficient technique for obtaining high-quality cell-free extracts from Aspergillus ochraceus.
• This preparation method is suitable for the reliable investigation of various in vitro fungal enzyme systems.
• The preservation of enzyme activity makes these extracts valuable for biochemical and enzymatic research in mycology.