Isolation of glucan particles and uses thereof

Abstract

The present invention relates to the isolation of novel glucan particles but also to mannoprotein from natural sources such as yeast cell walls, novel isolation methods, and the use of products thereof.

1 . Glucan particles with a molecular weight (M W  or M n ) of more than 100 000 after solubilization by carboxymethylation having preserved porous and fibrous structural properties.  
     
     
         2 . Glucan particles according to  claim 1  with a size range of 0.1 to 25 micrometers, preferably 0.5 to 15 micrometers, and most preferably 2 to 10 micrometers.  
     
     
         3 . Glucan particles according to claims  1  or  2 , which are insoluble in water and in most of the known organic solvents, having activated pores and which are showing an increased stability at high pH-values.  
     
     
         4 . Glucan particles according to  claims 1  to  3  with gel forming properties.  
     
     
         5 . Glucan particles according to at least one of the  claims 1  to  4  forming a stable gel in organic solvents or acidified water, when an aqueous suspension of these particles is heated to a temperature higher than 80° C.  
     
     
         6 . Process for isolating glucan particles from unicellular organisms or glucan-containing plants, comprising the steps 
 a) extracting mannoproteins with water at temperatures above the boiling point of water and/or 
 treatment of the raw materials or of the product from step a)  
   b) with proteases or 
 by non-denaturating chemical means for the removal of proteins and/or  
   c) with lipases, or 
 by solvent extraction for the removal of lipids on condition that steps b) and c) may be proceeded in any sequence and if necessary in dependence on the starting material one or two steps may be left out.  
   
     
     
         7 . Process according to  claim 6  comprising as step a) treatment with water at temperatures above the boiling point of water under elevated pressure and adjusted pH for extracting mannoprotein.  
     
     
         8 . Process according to at least one of the claims  6  or  7  wherein in step a) mannoproteins are extracted from an aqueous cell wall suspension in the concentration range of 1-20% by weight, preferably 10-20%, most preferably 13-17%, with adjusted pH in the range of 5-9, preferably pH 6-8, most preferably pH 6.5-7.5, at a temperature in the range of 120-130° C. for a defined period of time in the range of 0.1-24 hours, preferably 1-10 hours, most preferably 3-7 hours under elevated pressure.  
     
     
         9 . Process according to at least one of the claims  6 - 8 , wherein the mannoprotein is removed from the recovered aqueous solution (water extracts) from step a) by adding and mixing with an alcohol and precipitating.  
     
     
         10 . Process according to  claim 9  wherein the alcohol is selected from the group methanol, ethanol, propanol, and butanol.  
     
     
         11 . Process according to  claim 6  comprising as step b) treatment with proteases after pH adjustment at optimal level for the removal of contaminating proteins.  
     
     
         12 . Process according to  claim 6  wherein in step b) protein contaminants are extracted either with aqueous alkaline solution selected from the group sodium carbonate and sodium hydrogen carbonate at low concentration and low temperature or 
 with sodium dodecylsulfate at a concentration in the range of 0.1-5% by weight, preferably 1-3% by weight, most preferably 1.5-2.5% by weight.  
 
     
     
         13 . Process according to  claim 6  comprising as step c) treatment with lipases after pH adjustment at optimal level for removal of contaminating lipids.  
     
     
         14 . Process according to  claim 6  wherein in step c) lipid contaminants are extracted either with organic solvents, that are miscible with water, selected from the group acetone ethanol, methanol, isopropanol and butanol or mixtures thereof; or 
 with organic solvents that are not miscible with water, selected from the group isobutylketone, hexane, chloroform, methylenechloride, tetrachloroethylene and ethylacetate or mixtures thereof, or  
 with mixtures of organic solvents that are miscible with water with organic solvents that are not miscible with water; or  
 extraction with supercritical CO 2 , or  
 extraction with supercritical CO 2  and organic solvents as modifiers.  
 
     
     
         15 . Process according to claims  6  and  14  wherein in step c) lipid contaminants are extracted with a mixture of methanol and chloroform in a ratio of volume of 1:1 or of hexane and isopropanol in a ratio of volume of 3:2.  
     
     
         16 . Mannoprotein obtained from step a) according to at least one of the claims  6 - 10 .  
     
     
         17 . Glucan particles obtainable by a process according to at least one of the claims  6 - 8 ,  11 - 15 .  
     
     
         18 . Glucan particles obtainable by a process according to claims  6 - 8  by protease treatment after extracting mannoproteins with water.  
     
     
         19 . Glucan particles obtainable by a process according to at least one of the claims  6 - 8 ,  11 ,  13 , by lipase treatment or solvent extraction after extracting mannoprotein with water and removing protein contaminants with protease.  
     
     
         20 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  as adsorbents and carriers, as chromatographic active material or as adsorbents for toxic environmental compounds.  
     
     
         21 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  and/or mannoprotein according to  claim 16  for the preparation of pharmaceutical or cosmetic formulations or as additive in food and feed or in products needed in agriculture in crop protection.  
     
     
         22 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  and/or mannoprotein according to  claim 16  for the preparation of pharmaceutical formulations with immune system activating properties.  
     
     
         23 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  and/or mannoprotein according to  claim 16  for the preparation of pharmaceutical formulations with antitumor activity or for administration in combination with chemotherapy or dialysis.  
     
     
         24 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  and/or mannoprotein according to  claim 16  for the preparation of pharmaceutical formulations to improve the host-defences to bacterial or virus infections.  
     
     
         25 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  and/or mannoprotein according to  claim 16  for the preparation of pharmaceutical formulations with prophylactic activity against diseases of age.  
     
     
         26 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  and/or mannoprotein according to  claim 16  for the preparation of formulations with cholesterol reduction activity.  
     
     
         27 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  and/or mannoprotein according to  claim 16  for the preparation of formulations with an glucose regulating effect.  
     
     
         28 . Use of glucan particles according to at least one of the claims  1 - 5 ,  17 - 19  or obtainable from a process according to at least one of the claims  6 - 8  or  11 - 15  and/or mannoprotein according to  claim 16  for the preparation of formulations with improving effects on cardiovascular diseases, on autoimmune conditions like HIV, arthritis, lupus, allergic asthma or multipe sclerosis.