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10/06/2008
General Properties of P-Mapa
P-MAPA is a non-linear, high-molecular weight biopolymer, defined as a protein magnesium ammonium phospholinoleate-palmitoleate-anhydride polymer aggregated with immunomodulating properties
Macroscopic appearance of P-MAPA
• Chemical Name
Protein magnesium ammonium phospholinoleate-palmitoleate-anhydride polymer aggregated
• Abbreviated Name
P-MAPA or MAPA
• Chemical Analysis
Qualitative analyses indicated that the main components of P-MAPA are Mg+2 , NH4+, phosphate, linoleic acid, palmitoleic acid and protein.
Quantitative analysis was the following:
a) Protein: The Coomassie brilliant blue G-250 method was used to determine the protein content of P-M.A.P.A. Analysis gave a value of 0.49 ± 0.07% using bovine albumin as standard.
Molecular weight (protein) determined by gel filtration on Sephacryl-S-3000 was near of 10 kDa.
The content of amino acid were distributed (g/100 g protein) as follows: ASP (7.19%),THR(3.56%), SER (7.56%), GLU (8.53%), PRO (0.5%), GLY (9.69%), ALA (7.476), VAL (1.0%), MRT 4.38%), ISOLEU (2.54%), LEU (3.03%) TYR (0.5%), PHE (1.0%) HIS (2.73%), LYS (3.56%), TRP (1.3%) and ARG (35.2%).
The elemental analysis gave: 38.9% C; 6.1% H; 25.9%; 28.4% N and 0.6% S
Empirical formula: (C18H35Mg2NO21P5)391 (C326H614O163N204S2)0.16
b) Phosphate: Phosphate was determined spectrophotometrically using the ammonim molibdate/Zinc acetate, pH 5.0, method ( 350 nm, E= 7200 M-1 cm-1,. The phosphate content was 45.2 ± 2.7%
c) Magnesium: The atomic absorption method for quantification of magnesium was applied. Magnesium content was 20.1 ± 0.9%.
The fluorescence method using 8-hydroxyquinolein sulfonate was carried out at excitation = 420 nm
d) Lipids: The total lipid content was 11.6 ± 4.3%). The analysis showed 11,6 ± 4,3% of total lipids (22,7 ± 5,0% of palmitoleic acid, 42,9 ± 2,0% of linoleic acid and 32,0 ± 3,0% of oxidized linoleic acid.
e) Ammonium: The ammonium content was 10.0 ± 3.3%.
Molecular weight (determined by gel filtration on Sephacryl-S-3000): ~320 kDa
Crystals of P-MAPA ( 50 μm)
Images by S.E.M. (Scanning Electron Microscopy)
Nanocrystals of P-MAPA (400 nm )
Images by S.E.M. (Scaning Electron Microscopy)