Abstract: The presently disclosed subject matter relates to feed additive formulations for monogastric animal feed. Particularly, the disclosed formulations comprise an isolated xylanase enzyme and a B. licheniformis strain PWD-1. The feed additive formulations may further include B. amyloliquefaciens strain Ba-BPD1. The disclosed formulations are useful for addition to feeds for monogastric animals to synergistically improve the performance of the animals.

Abstract: Proteolytic enzyme variants that exhibit improved thermal stability relative to the wild type protease Bacillus licheniformis PWD-1 protease kerA are provided. The protease variants are derived by the mutation of at least one amino acid residue of the mature Bacillus licheniformis PWD-1 protease kerA. The variant proteases are useful as animal feed additives to improve the digestibility and/or the nutritional value of the animal feed.

Abstract: Methods and kits are provided for measuring protease activity in samples such as feed or food samples. The methods include adding a water insoluble substrate with a signal producing group to a feed or food sample containing the protease activity to be measured, incubating the sample in phosphate-free buffer such that the signal is produced, and measuring the amount of protease activity in the sample. The methods do not require separation of the incubation buffer from the feed or food and allow for visual inspection of a colorimetric signal for a semi-quantitative measurement of the in-feed/in-food protease activity. Thus, the assay is advantageous as it can be performed on-site without the use of laboratory equipment.

Abstract: Methods and kits are provided for measuring protease activity in samples such as feed or food samples. The methods include adding a water insoluble substrate with a signal producing group to a feed or food sample containing the protease activity to be measured, incubating the sample in phosphate-free buffer such that the signal is produced, and measuring the amount of protease activity in the sample. The methods do not require separation of the incubation buffer from the feed or food and allow for visual inspection of a colorimetric signal for a semi-quantitative measurement of the in-feed/in-food protease activity. Thus, the assay is advantageous as it can be performed on-site without the use of laboratory equipment.

Abstract: The present invention provides methods for reducing the production of nitric oxide, inducible nitric oxide synthase protein and mRNA, and cyclooxygenase-2 protein and mRNA in cells both in vivo and in vitro via the administration of extracts and compounds derived from the seeds of Pycnanthus angolensis Warb. (P. Kombo), commonly known as African nutmeg. These extracts and compounds, namely, kombo butter, kombo butter acid extract, sargaquinoic acid, sargachromenol, and sargahydroquinoic acid, are also useful in the treatment and prevention of a battery of adverse health conditions in human, animal, and avian subjects that are advanced by the production of nitric oxide or its metabolites and/or by the activity of cyclooxygenase-2. Methods for the isolation of kombo butter acid extracts are also provided.

Abstract: A method and composition for destruction of infectious prion proteins in tissue, by thermal/enzymatic treatment of the tissue with a prion-destructive protease. The method and composition are applicable to treatment of tissue containing or contaminated with prion protein strains associated with transmissible spongiform encephalopathy (TSE) and/or other prion protein-mediated diseases.

Abstract: A method and composition for destruction of infectious prion proteins in tissue, by thermal/enzymatic treatment of the tissue with a prion-destructive protease. The method and composition are applicable to treatment of tissue containing or contaminated with prion protein strains associated with transmissible spongiform encephalopathy (TSE) and/or other prion protein-mediated diseases.

Abstract: A method for purifying a plant protein comprises: (a) preparing a crude plant extract; (b) passing the crude plant extract with a first buffer solution through a guard column comprising a hydrophobic interaction chromatography medium of hydrophobicity sufficiently high to prevent tannins and polyphenols from eluting from the column in the presence of the buffer solution, but sufficiently low that a protein fraction elutes with the first buffer solution; (c) passing the protein fraction through a capture column coupled in series to the guard column, the capture column comprising a hydrophobic interaction chromatography medium of hydrophobicity sufficiently high to prevent the protein from eluting in the presence of the buffer solution; and (d) eluting the protein from the capture column as a purified fraction. Preferably the plant is miracle fruit, the protein is Miraculin, and the method comprises ion exchange chromatography and gel filtration chromatography of the purified fraction of the protein.