Plant mangrove-associated fungus Culvularia lunata and a simple and efficient method of obtaining high yield of pure mannitol from the same

Abstract

The present invention relates to a plant mangrove-associated fungus Culvularia lunata of International deposition no. ______, and a simple and efficient method of obtaining high yield of pure mannitol from a plant mangrove-associated fungus Curvularia lunata , said method comprising steps of cutting the leaves into small pieces and placing it on potato dextrose agar (PDA) plates for 48 hours, maintaining the culture at temperature ranging between 26-47° C. and salinity ranging between 4-32 ppt with occasional stress conditions for about 17-19 days to obtain mycelial mat, sonicating the mat to lyse the cells, extracting crude from the sonicated mat using methanol repeatedly, concentrating the crude, treating the concentrated crude extract with solvents of increasing polarity, obtaining aqueous fraction as white powder residue after the treatment containing crude mannitol, purifying the crude mannitol by chromatography to obtain pure mannitol with about 75% of the total crude extract.

1 . A plant mangrove-associated fungus  Culvularia lunata  of International deposition no. ______.  
     
     
         2 . The fungus as claimed in  claim 1 , wherein the mangrove plant is  Acanthus illicifolius.    
     
     
         3 . The fungus as claimed in  claim 1 , wherein the fungus is fast growing fungus.  
     
     
         4 . The fungus as claimed in  claim 1 , wherein the fungus is of color brown to blackish brown with black reverse.  
     
     
         5 . The fungus as claimed in  claim 1 , wherein the fungus has conidia of pale brown color with three or more traverse septa and are formed apically through a pore (poroconidia) in a sympodially elongated geniculate conidiophore.  
     
     
         6 . The fungus as claimed in  claim 1 , wherein the fungus shows conidia of cylindrical or slightly curve shape, with one of the central cells being larger and darker.  
     
     
         7 . A simple and efficient method of obtaining high yield of pure mannitol from a plant mangrove-associated fungus  Curvularia lunata  of International deposition No. ______, said method comprising steps of: 
 a. obtaining leaves from the plant mangrove,    b. rinsing the leaves in sterile seawater and keeping them in sterile, moist culture chamber for about two weeks,    c. cutting the leaves into small pieces and placing it on potato dextrose agar (PDA) plates for about 48 hours,    d. transferring individual colonies aseptically into a sterile fresh PDA plate to obtain pure culture,    e. maintaining the isolated fungal cultures on PDA slants for about 7 days to obtain mycelia,    f. transferring the mycelia to a fresh PDB with seawater and distilled water using starch and sugar as source of carbon.    g. incubating the culture at temperature ranging between 26-32° C. for about 3-6 days to obtain inoculum,    h. inoculating the PBD having seawater and distilled with the inoculum,    i. maintaining the culture at temperature ranging between 26-47° C. and salinity ranging between 4-32 ppt with occasional stress conditions for about 17-19 days to obtain mycelial mat,    j. sonicating the mat to lyse the cells,    k. extracting crude from the sonicated mat using methanol repeatedly,    l. concentrating the crude,    m. treating the concentrated crude extract with solvents of increasing polarity,    n. obtaining aqueous fraction as white powder residue after the treatment containing crude mannitol,    o. purifying the crude mannitol by chromatography to obtain pure mannitol with about 75% of the total crude extract.    
     
     
         8 . A method as claimed in  claim 7 , wherein the plant is  Acanthus illicifolius.    
     
     
         9 . A method as claimed in  claim 7 , wherein the starch is of concentration ranging between 3-5 g/l.  
     
     
         10 . A method as claimed in  claim 7 , wherein the dextrose is of the concentration ranging between 18-22 g/l.  
     
     
         11 . A method as claimed in  claim 7 , wherein the ratio of seawater and distilled water is ranging between 1:5 to 5:1, preferably 1:1.  
     
     
         12 . A method as claimed in  claim 7 , wherein the stress conditions is temperature ranging between 40-45° C.  
     
     
         13 . A method as claimed in  claim 7 , wherein the stress conditions is salinity ranging between 15-17 ppt.  
     
     
         14 . A method as claimed in  claim 7 , wherein the stress conditions lead to high percentage yield of mannitol.  
     
     
         15 . A method as claimed in  claim 7 , wherein the leaves are young, and living leaves.  
     
     
         16 . A method as claimed in  claim 7 , wherein concentrating the crude using vacuum evaporator at temperature ranging between 28-32° C.  
     
     
         17 . A method as claimed in  claim 7 , wherein the solvents in increasing polarity are petroleum ether, chloroform, and ethyl acetate.  
     
     
         18 . A method as claimed in  claim 7 , wherein the chromatography is on G-10 solid support.