title: Simple, effective protein extraction method and proteomics analysis from polyunsaturated fatty acids-producing micro-organisms.
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.
authors.affiliation: Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China. xpling@xmu.edu.cn.|The Key Lab for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen, 361005, China. xpling@xmu.edu.cn.
authors.forename: Xueping
authors.initials: X
authors.lastname: Ling
authors.affiliation: Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Guo
authors.affiliation: Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
authors.forename: Chuqiang
authors.initials: C
authors.lastname: Zheng
authors.affiliation: Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
authors.forename: Chiming
authors.initials: C
authors.lastname: Ye
authors.affiliation: Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.|The Key Lab for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen, 361005, China.
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
authors.forename: Xueshan
authors.initials: X
authors.lastname: Pan
authors.affiliation: Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
authors.forename: Zhengqi
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China. yswu@mail.ncku.edu.tw.|Department of Chemical Engineering, National Cheng Kung University, Tainan, 70101, Taiwan. yswu@mail.ncku.edu.tw.|Research Center for Energy Technology and Strategy, National Cheng Kung University, Tainan, 70101, Taiwan. yswu@mail.ncku.edu.tw.
authors.forename: I-Son
authors.initials: IS
authors.lastname: Ng
mesh_terms: D018533:Biomass; D020093:Cunninghamella / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis*; D005285:Fermentation; D005656:Fungal Proteins / Q000302:isolation & purification*; D050356:Lipid Metabolism; D020094:Mortierella / Q000378:metabolism*; D040901:Proteomics; D058009:Stramenopiles / Q000378:metabolism*
chemical_list: D005231:Fatty Acids, Unsaturated; D005656:Fungal Proteins
keywords: Cold shock; Comparative proteomics; Oleaginous micro-organisms; Polyunsaturated fatty acids; Protein extraction