Compositions and Methods for Producing Fermentation Products and Residuals

Abstract

The present invention provides compositions and methods designed to increase value output of a fermentation reaction. In particular, the present invention provides a business method of increasing value output of a fermentation plant. The present invention also provides a modified fermentation residual of higher commercial value. Also provided in the present invention are complete animal feeds, nutritional supplements comprising the subject ferment residuals. Further provided by the present invention is a method of performing fermentation, a modified fermentative microorganism and a genetic vehicle for modifying such microorganism.

1 .- 37 . (canceled)  
   
   
       38 . A method of fermentation using carbon-containing material, comprising: 
 (a) mixing a carbon-containing material with a culture comprising genetically modified microorganisms that, in a fermentation reaction, produce an alcohol and a fermentation residual comprising a nutrient selected from the group consisting of amino acids, cofactors, hormones, proteins, vitamins and lipids, wherein the content of the nutrient in the fermentation residual is greater than that of an unmodified corresponding microorganism when used in the fermentation reaction;    (b) fermenting the culture under conditions suitable for production of the alcohol and under conditions suitable for production the nutrient;    (c) separating the alcohol from the culture; and    (d) producing the fermentation residual comprising the nutrient.    
   
   
       39 . The method of  claim 38 , wherein the microorganisms comprise a recombinant expression vector comprising an exogenous nucleotide sequence encoding a polypeptide and a regulatory sequence that controls the expression of the exogenous polypeptide, wherein expression of the exogenous polypeptide results in increased nutritional content of the fermentation residual compared with that of the unmodified microorganism.  
   
   
       40 . The method of  claim 39 , wherein the nutrient is an essential amino acid to at least one domesticated animal and the exogenous polypeptide comprises the essential amino acid.  
   
   
       41 . The method of  claim 40 , wherein the essential amino acid is selected from the group consisting of lysine, methionine, phenylalanine, threonine, isoleucine, tryptophan, valine, leucine, arginine, taurine and histidine.  
   
   
       42 . The method of  claim 39 , wherein the expression of the exogenous sequence is under the control of a regulatory sequence selected from the group consisting of a regulatory sequence of a heat shock gene, a regulatory sequence of a toxicity gene and a regulatory sequence of a spore formation gene.  
   
   
       43 . The method of  claim 38 , wherein the genetic modification modifies at least one of the structural genes in the nutrient's synthetic pathway.  
   
   
       44 . The method of  claim 38 , wherein the genetic modification modifies a regulatory control of the nutrient's synthetic pathway.  
   
   
       45 . The method of  claim 44 , wherein the synthetic pathway is for an essential amino acid for a domesticated animal.  
   
   
       46 . The method of  claim 38 , wherein the genetic modification modifies a structural gene that regulates synthesis of a peptide containing at least one essential amino acid for a domesticated animal.  
   
   
       47 . The method of  claim 38 , wherein the genetic modification modifies the nutrient's transport processes out of or into the microorganism.  
   
   
       48 . The method of  claim 39 , wherein expression of the exogenous sequence is induced when the fermentation reaction has achieved at least about 50% completion.  
   
   
       49 . The method of  claim 48 , wherein the at least about 50% completion is evidenced by a decrease in glucose content to less than about 50% of an initial content of glucose present in a fermentation reaction mixture prior to beginning the fermentation reaction.  
   
   
       50 . The method of  claim 39 , wherein expression of the exogenous nucleotide sequence depends on glucose concentration.  
   
   
       51 . The method of  claim 38 , wherein the nutrient is an essential amino acid to at least one domesticated animal.  
   
   
       52 . The method of  claim 51 , wherein the essential amino acid is selected from the group consisting of lysine, methionine, phenylalanine, threonine, isoleucine, tryptophan, valine, leucine, arginine, taurine and histidine.  
   
   
       53 . The method of  claim 38 , wherein the nutrient is a vitamin.  
   
   
       54 . The method of  claim 53 , wherein the vitamin is selected from the group consisting of vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin C, vitamin D1-D4, a tocopherol, and vitamin K.  
   
   
       55 . The method of  claim 38 , wherein the nutrient is a lipid.  
   
   
       56 . The method of  claim 38 , wherein the alcohol is ethanol.  
   
   
       57 . The method of  claim 38 , wherein the alcohol is selected from the group consisting of methanol, propanol and butanol.  
   
   
       58 . The method of  claim 38 , wherein the carbon-containing material is selected from the group consisting of cellulose, wood chips, vegetables, biomass, excreta, animal wastes, oat, wheat, corn, barley, milo, millet, rice, rye, sorghum, potato, sugar beets, taro, cassava, fruits, fruit juices, and sugar cane.  
   
   
       59 . The method of  claim 38 , wherein the fermentation residual comprises distillers dried grains.  
   
   
       60 . The method of  claim 38 , wherein the fermentation residual comprises distillers dried grains with solubles.  
   
   
       61 . The method of  claim 38 , wherein comprising incorporating the fermentation residual into animal feed.  
   
   
       62 . The method of  claim 38 , wherein the alcohol is separated by distillation.  
   
   
       63 . The method of  claim 38 , further comprising mixing the alcohol with another fuel.  
   
   
       64 . The method of  claim 38 , wherein the nutrient is produced when fermentation has substantially been completed.  
   
   
       65 . The method of  claim 38 , wherein the microorganism is yeast.  
   
   
       66 . The method of  claim 65 , wherein the yeast is a  Saccharomyces.    
   
   
       67 . The method of  claim 38 , wherein the microorganism is  Clostridium.    
   
   
       68 . The method of  claim 38 , wherein the microorganism is selected from the group consisting of Zymomonas sp.,  E. coli, Corynebacterium, Brevibacterium  and  Bacillus  ssp.  
   
   
       69 . A genetically modified microorganism that, in a fermentation reaction, produces an alcohol and a fermentation residual comprising a nutrient selected from the group consisting of amino acids, cofactors, hormones, proteins, vitamins, and lipids, wherein the content of the nutrient in the fermentation residual is greater than that of an unmodified corresponding microorganism when used in the fermentation reaction.  
   
   
       70 . The genetically modified microorganism of  claim 69 , comprising a recombinant expression vector comprising an exogenous nucleotide sequence encoding a polypeptide and a regulatory sequence that controls the expression of the exogenous polypeptide, wherein expression of the exogenous polypeptide results in increased nutritional content of the fermentation residual compared with that of the unmodified microorganism.  
   
   
       71 . The genetically modified microorganism of  claim 70 , wherein the nutrient is an essential amino acid to at least one domesticated animal and the exogenous polypeptide comprises the essential amino acid.  
   
   
       72 . The genetically modified microorganism of  claim 71 , wherein the essential amino acid is selected from the group consisting of lysine, methionine, phenylalanine, threonine, isoleucine, tryptophan, valine, leucine, arginine, taurine and histidine.  
   
   
       73 . The genetically modified microorganism of  claim 70 , wherein the expression of the exogenous sequence is under the control of a regulatory sequence selected from the group consisting of a regulatory sequence of a heat shock gene, a regulatory sequence of a toxicity gene and a regulatory sequence of a spore formation gene.  
   
   
       74 . The genetically modified microorganism of  claim 69 , wherein the genetic modification modifies at least one of the structural genes in the nutrient's synthetic pathway.  
   
   
       75 . The genetically modified microorganism of  claim 69 , wherein the synthetic pathway is for an essential amino acid for a domesticated animal.  
   
   
       76 . The genetically modified microorganism of  claim 69 , wherein the genetic modification modifies a regulatory control of the nutrient's synthetic pathway.  
   
   
       77 . The genetically modified microorganism of  claim 69 , wherein the genetic modification modifies a structural gene that regulates synthesis of a peptide containing at least one essential amino acid for a domesticated animal.  
   
   
       78 . The genetically modified microorganism of  claim 69 , wherein the genetic modification modifies the nutrient's transport processes out of and into the microorganism.  
   
   
       79 . The genetically modified microorganism of  claim 70 , wherein expression of the exogenous sequence is induced when the fermentation reaction has achieved at least about 50% completion.  
   
   
       80 . The genetically modified microorganism of  claim 79 , wherein the at least about 50% completion is evidenced by a decrease in glucose content to less than about 50% of an initial content of glucose present in a fermentation reaction mixture prior to beginning the fermentation reaction.  
   
   
       81 . The genetically modified microorganism of  claim 69 , wherein expression of the exogenous nucleotide sequence depends on glucose concentration.  
   
   
       82 . The genetically modified microorganism of  claim 69 , wherein the nutrient is an essential amino acid to at least one domesticated animal.  
   
   
       83 . The genetically modified microorganism of  claim 82 , wherein the essential amino acid is selected from the group consisting of, lysine, methionine, phenylalanine, threonine, isoleucine, tryptophan, valine, leucine, arginine, taurine and histidine.  
   
   
       84 . The genetically modified microorganism of  claim 69 , wherein the nutrient is a vitamin.  
   
   
       85 . The genetically modified microorganism of  claim 84 , wherein the vitamin is selected from the group consisting of vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin C, vitamin D1-D4, a tocopherol, and vitamin K.  
   
   
       86 . The genetically modified microorganism of  claim 69 , wherein the nutrient is a lipid.  
   
   
       87 . The genetically modified microorganism of  claim 69 , wherein the alcohol is ethanol.  
   
   
       88 . The genetically modified microorganism of  claim 69 , wherein the alcohol is selected from the group consisting of methanol, propanol, and butanol.  
   
   
       89 . The genetically modified microorganism of  claim 69 , wherein the microorganism is yeast.  
   
   
       90 . The genetically modified microorganism of  claim 89 , wherein the yeast is a  Saccharomyces.    
   
   
       91 . The genetically modified microorganism of  claim 69 , wherein the microorganism is  Clostridium.    
   
   
       92 . The genetically modified microorganism of  claim 69 , wherein the microorganism is selected from the group consisting of  Zymomonas  sp.,  E. coli, Corynebacterium, Brevibacterium  and  Bacillus  ssp.  
   
   
       93 . A fermentation culture comprising: 
 (a) a genetically modified microorganism that, in a fermentation reaction, produces an alcohol and a fermentation residual comprising a nutrient selected from the group consisting of an amino acid, a cofactor, a hormone, a protein, a vitamin, and a lipid, wherein the content of the nutrient in the fermentation residual is greater than that of an unmodified corresponding microorganism when used in the fermentation reaction and    (b) a fermentation medium for production of nutrients comprising a carbon source.    
   
   
       94 . The culture of  claim 93  wherein the carbon source is selected from cellulose, wood chips, vegetables, biomass, excreta, animal wastes, oat, wheat, corn, barley, milo, millet, rice, rye, sorghum, potato, sugar beets, taro, cassava, fruits, fruit juices, and sugar cane.  
   
   
       95 . The culture of  claim 93 , wherein the alcohol is ethanol.  
   
   
       96 . The culture of  claim 93 , wherein the alcohol is selected from the group consisting of methanol, propanol, and butanol.  
   
   
       97 . The culture of  claim 93 , wherein the microorganism is yeast.  
   
   
       98 . The culture of  claim 97 , wherein the yeast is a  Saccharomyces.    
   
   
       99 . The culture of  claim 93 , wherein the microorganism is  Clostridium.    
   
   
       100 . The culture of  claim 93 , wherein the microorganism is selected from the group consisting of Zymomonas sp.,  E. coli, Corynebacterium, Brevibacterium  and  Bacillus  ssp.  
   
   
       101 . An expression vector comprising an exogenous sequence encoding a polypeptide comprising at least one essential amino acid for a domesticated animal, wherein expression of the exogenous sequence is induced when a fermentation reaction producing an alcohol or an alkane has achieved at least about 50% completion.  
   
   
       102 . The expression vector of  claim 101 , wherein expression of the exogenous sequence is under the control of a regulatory sequence selected from the group consisting of a glucose suppressor operon, regulatory sequence of a heat shock gene, regulatory sequence of a toxicity gene, regulatory sequence of a spore formation gene.  
   
   
       103 . The expression vector of  claim 101 , wherein at least about 5% of the amino acid residues contained in the polypeptide are essential amino acids for a domesticated animal.