Bile resistant bacillus composition

Abstract

A bacillus composition characterized by fast germination and outgrowth in bile salts (simulated gut environment) and by producing a compound of interest. The bacillus composition may be used as supplement in animal feed where it has a probiotic (health promoting) effect and increases the digestion and availability of nutrients from animal feeds.

1 - 12 . (canceled) 
     
     
         13 . A  Bacillus  composition comprising from 10 5  to 10 12  CFU/g  Bacillus  spore cells of a mutant  Bacillus  strain, wherein:
 (a) the  Bacillus  spores have a germination and outgrowth from spore to vegetative cell in the presence of a bile salt medium comprising 4 mM or 6 mM bile salt such that the spores reach a vegetative cell growth point of 0.4 OD630 within less than 18 hours or 19 hours, respectively; and   (b) the  Bacillus  vegetative cells overproduce at least one desired compound in comparison to the wild-type  Bacillus  strain from which the mutant  Bacillus  strain derives when grown under comparable conditions.   
     
     
         14 . The  Bacillus  composition of  claim 13 , wherein the desired compound is an enzyme. 
     
     
         15 . The  Bacillus  composition of  claim 14 , wherein the enzyme is phytase or mannanase. 
     
     
         16 . The  Bacillus  composition of  claim 13 , wherein the desired compound is an amino acid. 
     
     
         17 . The  Bacillus  composition of  claim 16 , wherein the amino acid is selected from the group consisting of phenylalanine, valine, threonine, tryptophan, isoleucine, methionine, leucine, lysine, cysteine, tyrosine, histidine, and arginine. 
     
     
         18 . The  Bacillus  composition of  claim 13 , wherein the  Bacillus  spore cells are present as dried spore cells. 
     
     
         19 . The  Bacillus  composition of  claim 13 , wherein the  Bacillus  is  Bacillus subtilis.   
     
     
         20 . A method for isolating a mutant  Bacillus  strain which germinates and outgrows rapidly in the presence of a bile salt and which overproduces at least one desired compound, comprising:
 (a) subjecting a wild-type  Bacillus  spore cell to mutagenesis to isolate a  Bacillus  spore cell having a germination and outgrowth from spore to vegetative cell in the presence of a bile salt medium comprising 4 mM or 6 mM bile salt such that the  Bacillus  spores reach a vegetative cell growth point of 0.4 OD630 within less than 18 hours or 19 hours;   (b) subjecting vegetative cells grown from the isolated spore cell of step (a) to mutagenesis, thereby obtaining a pool of individual vegetative cells which retain rapid germination and outgrowth in the presence of a bile salt; and   (c) selecting a vegetative cell from the pool of step (b), which vegetative cell retains rapid germination and outgrowth in the presence of a bile salt, and which overproduces the at least one desired compound.   
     
     
         21 . The method of  claim 20 , wherein the  Bacillus  strain is  Bacillus subtilis.   
     
     
         22 . A method for feeding an animal comprising administering the  Bacillus  composition of claim  1  to the animal.