[Purification and properties of Se-containing allophycocyanins from selenium rich Spirulina platensis].
Three Se-containing allophycocyanins (Se-APC) with high purity were purified from Se rich Spirulina platensis (Se-sp.) by hydroxyapatite chromatography, DEAE-52 anion-exchange chromatography and native gel preparative electrophoresis. Their biochemicial properties were explored by spectral scanning and electrophoresis analysis of Native-PAGE, SDS-PAGE and IEF on thin slab gel. Protein molecular weight (MW) of APC aggregation was determined by gel filter on Sephadex G-200 column. Se content of native and denatured Se-APC was detected by 2, 3-DAN fluorocence method. According to visible and fluorescence spectral character, three purified fractions of APC were identified to be APCI, APCII and APCIII. Native-PAGE and SDS-PAGE analysis revealed that they all shaped trimer (alphabeta) 3 of alpha and beta subunit with molecular mass of 18.3kDa and 15.7kDa, whereas APCI contains gamma subunit (about 32kDa) visibly and APCIII maybe contain the linker peptide of L(C)(8 - 10 kDa) based on their MW to be determined of 130.9, 98.1 and 106.30 kDa. IEF detection showed that the pl of Se-APCs was 4.76, 4.85 and 5.02 respectively. Se content of three purified Se-APCs were 316, 273 and 408 microg/g, which decreased about 25% after deaggregation treatment by 0.50 mol/L NaSCN and decreased more than 50% after denaturation treatment by 2-mercaptoethanol and reached to a steady content of 132 microg/g on average. These results indicated that Se incorporation into APC had no influence on function of energy transfer as well as biochemical property of APCs, and Se binding with APCs was highly relevant to its aggregation states whereas Se integrated steadily with its subunits.
Huang
Zhi
Z
College of Life Science and Technology, Jinan University, Guangzhou 510632, China. thsh@jnu.edu.cn
Yang
Fang
F
Zheng
Wen-Jie
WJ
D000042:Absorption; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010798:Phycocyanin / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D011489:Protein Denaturation; D055503:Protein Multimerization / Q000187:drug effects; D020836:Protein Structure, Quaternary; D012643:Selenium / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D053545:Spirulina / Q000737:chemistry / Q000378:metabolism*; D013861:Thiocyanates / Q000494:pharmacology
D001426:Bacterial Proteins; D013861:Thiocyanates; C002497:allophycocyanin; D010798:Phycocyanin; D012643:Selenium; C009941:potassium thiocyanate