Microbial cell disruption methods for efficient release of enzyme L-asparaginase

Area of Science:

• Biotechnology
• Microbiology
• Biochemistry

Background:

• Efficient cell disruption is crucial for extracting intracellular enzymes like L-asparaginase.
• Various methods exist for microbial cell lysis, but their effectiveness varies across different microorganisms.

Purpose of the Study:

• To evaluate and compare the efficacy of different laboratory-scale cell disruption methods for L-asparaginase extraction.
• To identify the optimal disruption procedure for specific microbial sources: Escherichia coli, Leucosporidium muscorum, and Aspergillus terreus.

Main Methods:

• Assessed cell disruption using glass bead stirring, sonication, osmotic shock, freezing/grinding, solvents, and detergents.
• Measured total protein release and L-asparaginase activity as indicators of disruption efficiency.
• Tested methods on Escherichia coli BL21 (DE3), Leucosporidium muscorum, and Aspergillus terreus (CCT 7693).

Main Results:

• Sonication and glass bead stirring were effective for filamentous fungi, with sonication yielding higher specific activity (4.61 IU/mg).
• Mechanical methods, especially sonication, were superior for yeast and bacteria, achieving higher total enzyme activity (7.3 IU for yeast, 61.7 IU for bacteria) and specific activity (9.01 IU/mg for bacteria).
• Mechanical methods generally proved most effective for all microbial cell types studied.

Conclusions:

• Mechanical cell disruption techniques, particularly sonication, are highly effective for extracting intracellular enzymes from diverse microbial sources.
• This study provides a comparative analysis of cell disruption methods for L-asparaginase extraction, applicable to other periplasmic enzyme extractions.