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Simple, effective protein extraction method and proteomics analysis from polyunsaturated fatty acids-producing micro-organisms

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Abstract

Polyunsaturated fatty acids (PUFAs) are valuable ingredients in the food and pharmaceutical products due to their beneficial influence on human health. Most studies paid attention on the production of PUFAs from oleaginous micro-organisms but seldom on the comparative proteomics of cells. In the study, three methods (i.e., cold shock, acetone precipitation and ethanol precipitation) for lipid removal from crude protein extracts were applied in different PUFAs-producing micro-organisms. Among the selective strains, Schizochytrium was used as an oleaginous strain with high lipid of 60.3 (w/w %) in biomass. The Mortierella alpina and Cunninghamella echinulata were chosen as the low-lipid-content strains with 25.8 (w/w %) and 21.8 (w/w %) of lipid in biomass, respectively. The cold shock resulted as the most effective method for lipid removed, thus obtained higher protein amount for Schizochytrium. Moreover, from the comparative proteomics for the three PUFAs-producing strains, it showed more significant proteins of up or down-regulation were explored under cold shock treatment. Therefore, the essential proteins (i.e., polyunsaturated fatty acid synthase) and regulating proteins were observed. In conclusion, this study provides a valuable and practical approach for analysis of high PUFAs-producing strains at the proteomics level, and would further accelerate the understanding of the metabolic flux in oleaginous micro-organisms.

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Acknowledgments

The authors are grateful to the National Natural Science Foundation of China (No. 41206115), the Natural Science Foundation of Fujian Province of China (No. 2013J01060) and the National High-Tech R&D Program of China (No. 2014AA021701).

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Authors and Affiliations

  1. Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China

    Xueping Ling, Jing Guo, Chuqiang Zheng, Chiming Ye, Yinghua Lu, Xueshan Pan, Zhengqi Chen & I-Son Ng

  2. The Key Lab for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen, 361005, China

    Xueping Ling & Yinghua Lu

  3. Department of Chemical Engineering, National Cheng Kung University, Tainan, 70101, Taiwan

    I-Son Ng

  4. Research Center for Energy Technology and Strategy, National Cheng Kung University, Tainan, 70101, Taiwan

    I-Son Ng

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  1. Xueping Ling
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  2. Jing Guo
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Correspondence to Xueping Ling or I-Son Ng.

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Ling, X., Guo, J., Zheng, C. et al. Simple, effective protein extraction method and proteomics analysis from polyunsaturated fatty acids-producing micro-organisms. Bioprocess Biosyst Eng 38, 2331–2341 (2015). https://doi.org/10.1007/s00449-015-1467-7

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  • DOI: https://doi.org/10.1007/s00449-015-1467-7

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