Preparation and use of biofilm-degrading, multiple-specificity, hydrolytic enzyme mixtures

Abstract

The present invention relates to isolated structures containing degradative enzymes produced from a marine organism. The enzymes produced are based on the carbon source upon which the marine organism is growing. The enzymes are found in structures that can be isolated such that the degradative enzymes are easily harvested.

1 . Bacterial strain Microbulbifer 2-40.  
     
     
         2 . An isolated structure from Microbulbifer 2-40 comprising at least one degradative enzyme.  
     
     
         3 . The isolated structure according to  claim 2 , wherein the at least one degradative enzyme degrades a polysaccharide selected from alginate, araban, carrageenan, cellulose, carboxymethylcellulose, chitin, glycogen, β-glucan, pectin, laminarin, pullulan, starch, xylan, or agar.  
     
     
         4 . The isolated structure according to  claim 3 , wherein the polysaccharide is alginate.  
     
     
         5 . The isolated structure according to  claim 3 , wherein the polysaccharide is cellulose.  
     
     
         6 . The isolated structure according to  claim 3 , wherein polysaccharide is chitin.  
     
     
         7 . The isolated structure according to  claim 3 , wherein the polysaccharide is agar.  
     
     
         8 . The isolated structure according to  claim 2 , wherein the at least one degradative enzyme comprises at least two degradative enzymes.  
     
     
         9 . The isolated structure according to  claim 8 , wherein the at least two degradative enzymes are an agarase and a chitinase.  
     
     
         10 . The isolated structure of  claim 2 , obtained from a process comprising: 
 1) growing Microbulbifer 2-40 to mid-log phase in a medium comprising a polysaccharide;    2) centrifuging the medium to obtain a supernatant;    3) filtering the supernatant; and    4) purifying the tubules.    
     
     
         11 . The isolated structure according to  claim 10 , wherein the polysaccharide is alginate, araban, carrageenan, cellulose, carboxymethylcellulose, chitin, glycogen, β-glucan, pectin, laminarin, pullulan, starch, xylan, or agar.  
     
     
         12 . The isolated structure according to  claim 10 , wherein the medium is centrifuged at about 6,000 rpm for about 10 minutes.  
     
     
         13 . The isolated structure according to  claim 10 , wherein the supernatant is filtered through a filter paper with a pore size of about 2 μm to about 10 μm.  
     
     
         14 . The isolated structure according to  claim 2 , wherein the isolated structure is an isolated degradosome, an isolated vesicle, an isolated tubule, or an isolated elaborated structure.  
     
     
         15 . The isolated structure according to  claim 2 , wherein the isolated structure is an isolated tubule.  
     
     
         16 . A degradative enzyme associated with an isolated structure of Microbulbifer 2-40.  
     
     
         17 . The degradative enzyme of  claim 16 , wherein the enzyme degrades a polysaccharide selected from alginate, araban, carrageenan, cellulose, carboxymethylcellulose, chitin, glycogen, β-glucan, pectin, laminarin, pullulan, starch, xylan, or agar.  
     
     
         18 . The degradative enzyme of  claim 17 , wherein the polysaccharide is alginate, araban, carrageenan, cellulose, carboxymethylcellulose, chitin, glycogen, β-glucan, pectin, laminarin, pullulan, starch, xylan, or agar.  
     
     
         19 . The degradative enzyme of  claim 17 , wherein the polysaccharide is alginate.  
     
     
         20 . The degradative enzyme of  claim 17 , wherein the polysaccharide is cellulose.  
     
     
         21 . The degradative enzyme of  claim 17 , wherein the polysaccharide is chitin.  
     
     
         22 . The degradative enzyme of  claim 17 , wherein the polysaccharide is agar.