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Simple, effective protein extraction method and proteomics analysis from polyunsaturated fatty acids-producing
Xueping Ling1,2, Jing Guo3, Chuqiang Zheng3
1Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China. xpling@xmu.edu.cn.
Bioprocess and Biosystems Engineering
|September 23, 2015
Summary
Cold shock effectively removes lipids from PUFA-producing microorganisms, enabling better proteomic analysis. This method reveals key proteins involved in polyunsaturated fatty acid synthesis and regulation in oleaginous strains.
Area of Science:
- Biotechnology
- Microbiology
- Proteomics
Background:
- Polyunsaturated fatty acids (PUFAs) are vital for food and pharmaceutical industries due to health benefits.
- Research has focused on PUFA production by microorganisms, but comparative cell proteomics is underexplored.
Purpose of the Study:
- To evaluate lipid removal methods for proteomic analysis in PUFA-producing microorganisms.
- To identify key proteins involved in PUFA synthesis and regulation under different conditions.
Main Methods:
- Applied cold shock, acetone precipitation, and ethanol precipitation for lipid removal from crude protein extracts.
- Utilized comparative proteomics to analyze protein expression in Schizochytrium, Mortierella alpina, and Cunninghamella echinulata.
- Quantified lipid content in selected oleaginous and low-lipid-content strains.
Main Results:
- Cold shock was the most effective lipid removal method, yielding higher protein yields, especially for Schizochytrium.
- Comparative proteomics identified significant up/down-regulation of proteins, including polyunsaturated fatty acid synthase, under cold shock.
- Schizochytrium showed high lipid content (60.3%), while M. alpina (25.8%) and C. echinulata (21.8%) had lower lipid content.
Conclusions:
- Cold shock is a practical and effective method for preparing PUFA-producing microorganisms for proteomic analysis.
- This approach enhances understanding of metabolic flux and protein regulation in oleaginous microorganisms for improved PUFA production.
