10/06/2008

Properties of P-Mapa

General Properties of P-Mapa

 

P-MAPA is a aggregated polymer of protein magnesium ammonium phospholinoleate-palmitoleate anhydride from Aspergillus oryzae which immunomodulating activities.

 

• Chemical Name

 

Aggregated polymer of protein magnesium ammonium phospholinoleate-palmitoleate anhydride.

 

• 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 or the formula of C326H614O163N204S2.

 

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%.

 

P-MAPA - Proposed structural formula

 

 

Molecular weight (determined by gel filtration on Sephacryl-S-3000): 320 kDa.

 

 

SEM of P-MAPA crystals in purest form