Compositions, methods and uses for a novel family of peptides

Abstract

The present invention includes compositions and methods for the characterization and use of novel peptide from Brevibacillus sp., and peptides related thereto, including an isolated and purified, heat stable, amino terminus-methylated, carboxy-terminus reduced peptide that have two or more D-amino acids used as, e.g., an antimicrobial or even a feed additive.

1 . A feed additive comprising: an isolated and purified, heat stable, amino terminus-methylated, carboxy-terminus reduced peptide comprising two or more D-amino acids isolated from  Brevibacillus  sp., wherein the peptide is selected from at least one of SEQ ID NOS: 6-20. 
     
     
         2 . The additive of  claim 1 , wherein the Vol is valine alcohol produced by reducing —COOH group of the C-terminal Valine to —CH 2 OH. 
     
     
         3 . The additive of  claim 1 , wherein the carboxy-terminus —COOH group of the C-terminal Valine is reduced to —CH 2 OH and confers protease resistance to the peptide. 
     
     
         4 . The additive of  claim 1 , wherein the peptide is stable at a pH of 1.0, at a pH 13.0, resistant to proteases or combinations thereof. 
     
     
         5 . (canceled) 
     
     
         6 . The additive of  claim 1 , wherein the peptide kills, gram positive bacteria, gram negative bacteria, fungi, protozoa or combinations thereof. 
     
     
         7 . (canceled) 
     
     
         8 . The additive of  claim 1 , wherein the peptide is added at between about 0.5 and about 100 ppm. 
     
     
         9 . The additive of  claim 1 , wherein the peptide is added at between about 6 and about 12 ppm. 
     
     
         10 . The additive of  claim 1 , wherein the peptide is added to a feed adapted for use by one or more of poultry, livestock, farm-raised fish, crabs, shrimp and fresh-water turtles. 
     
     
         11 . The feed additive of  claim 1 , wherein the additive is added to a cereal-based animal feed comprising: at least one cereal selected from barley, soya, wheat, triticale, rye and maize; and an isolated and purified, heat stable, amino terminus-methylated, carboxy-terminus reduced peptide comprising two or more D-amino acids isolated from  Brevibacillus  sp., wherein the peptide comprises SEQ ID NOS: 2-20. 
     
     
         12 - 13 . (canceled) 
     
     
         14 . A biologically pure culture of microorganism  Brevibacillus texasporus  deposit No. ATCC PTA-5854) that produces an antimicrobial peptide that is carboxy-terminus reduced heat stable, amino terminus-methylated peptide and comprises two or more D-amino acids, wherein the peptide is selected from at least one of SEQ ID NOS: 1-20. 
     
     
         15 . The microorganism of  claim 14 , wherein the microorganism is added to a feed. 
     
     
         16 . The microorganism of  claim 14 , wherein the microorganism is mixed with a feed for livestock selected from the group consisting of a milk replacer, a grower feed, a finisher feed, a pre-starter feed and a starter feed. 
     
     
         17 . A method for increasing body weight gain efficiency and feed efficiency in animals, comprising mixing the composition of  claim 1  with an animal feed. 
     
     
         18 . The method of  claim 17 , wherein the animal feed is adapted for feeding livestock selected from the group consisting of a cattle, a swine, a chicken, a horse, a turkey, a sheep, a goat, a farm-raised fish, crab, shrimp and a turtle. 
     
     
         19 . The method of  claim 17 , wherein the animal feed is adapted for feeding birds selected from the group consisting of chicken, turkey, duck, quail, Cornish hens, and pigeon. 
     
     
         20 . The method of  claim 17 , wherein said feed is selected from the group consisting of a cereal, soybean meal, isolated soybean protein, isolated soybean oil, isolated soybean fat, skimmed milk, fish meal, meat meal, bone meal, blood meal, blood plasma protein, whey, rice bran, wheat bran, a sweetener, a mineral, a vitamin, salt, and grass. 
     
     
         21 . The method of  claim 17 , wherein the daily dose of the peptide ranges from about 1 milligram to about 10 grams per kg body weight of the animal. 
     
     
         22 - 23 . (canceled) 
     
     
         24 . An isolated and purified nucleic acid having the sequence of the BT operon (SEQ ID NO.: 21), or portions thereof, that express proteins that produce a heat stable, amino terminus-methylated, carboxy-terminus reduced peptide comprising two or more D-amino acids. 
     
     
         25 . The isolated and purified nucleic acid of  claim 24 , wherein the nucleic acid encodes one or more polypeptide sequences for BT operon proteins (SEQ ID NOS.: 22 to 28) that comprise one or more enzymes used to make a heat stable, amino terminus-methylated, carboxy-terminus reduced peptide comprising two or more D-amino acids, wherein the peptide is selected from at least one of SEQ ID NOS: 1-20, and the nucleic acid has been transformed into a bacteria. 
     
     
         26 - 28 . (canceled) 
     
     
         29 . The microorganism of  claim 14 , wherein the organism is an isolated bacterial comprising  Brevibacillus texasporus  E58. 
     
     
         30 - 35 . (canceled)