Method of detecting phytase activity or protease activity

Patent metadata

Field	Value
Publication number	US-9896714-B2
Application number	US-201013518785-A
Country	US
Kind code	B2
Filing date	Dec 17, 2010
Priority date	Dec 23, 2009
Publication date	Feb 20, 2018
Grant date	Feb 20, 2018

Abstract

A method of detecting a phytase activity or a protease activity is described. The method comprises the steps of: (a) providing a composition comprising a phytate/protein complex in a liquid or a gel; wherein the phytate/protein complex provides a detectable property to the composition; (b) providing a sample that comprises or is suspected of comprising phytase activity and/or protease activity, wherein the phytase and/or protease activity is capable of causing a change in the detectable property of the composition; (c) contacting the composition with the sample; and (d) determining if there is a detectable change in detectable property of the composition.

First claim

The invention claimed is:

1. A method of detecting a phytase activity or a protease activity comprising the steps of:
providing a medium comprising a continuous phase and a disperse phase, wherein the disperse phase comprises:
(1) a first component, the first component being a phytate or a phytic acid, and
(2) a second component, the second component being a protein,
wherein the first component and the second component are held together by one or more intermolecular interactions, wherein the phase provides a detectable property of turbidity to the medium;
providing a sample that comprises one or more of the phytase activity or the protease activity, wherein the one or more of the phytase activity or the protease activity are capable of affecting the disperse phase to cause a change in the turbidity of the medium;
contacting the medium with the sample; and
determining that there is a detectable change in the turbidity of the medium to determine an amount of the phytase activity or the protease activity in the sample.

2. The method of claim 1 , wherein the protein in the disperse phase of the medium is selected from a group comprising caseins, soy proteins, rapeseed proteins, mustard proteins, porcine haemoglobins, N,N-dimethylated caseins, bovine beta-lactoglobulins, bovine serum albumins, swine serum albumins, lysozymes, or proteins modified with a chromophore.

3. The method of claim 1 , wherein the medium is between a pH of 1.8 and a pH of 8.5.

4. The method of claim 1 , wherein the method comprises comparing the change in the detectable property of the medium to a control.

5. The method of claim 1 , wherein the sample is a biological sample.

6. The method of claim 5 , wherein the biological sample is an enzyme preparation comprised of a fermentation broth, a feed or in an extract of the fermentation broth or the feed.

7. The method of claim 1 , wherein determining if there is a detectable change in the detectable property of the medium comprises one or more of a quantitative assay, a semi-quantitative assay or a qualitative assay.

8. A method of detecting a phytase activity comprising the steps of:
providing a medium comprising a continuous phase and a disperse phase, wherein the disperse phase comprises:
(1) a first component, the first component being a phytate or a phytic acid, and
(2) a second component, the second component being a protein, wherein the first component and the second component are held together by one or more intermolecular interactions, wherein the disperse phase provides a detectable property of turbidity to the medium;
providing a sample that comprises the phytase activity, wherein the phytase activity is capable of affecting the disperse phase to cause a change in the turbidity of the medium;
contacting the medium with the sample, wherein one or more protease inhibitors are added to the sample in a sufficient amount as to inhibit any protease activity; and
determining that there is a detectable change in the turbidity of the medium to determine an amount of the phytase activity in the sample.

9. The method of claim 8 , wherein the protein in the disperse phase of the medium is selected from a group comprising caseins, soy proteins, rapeseed proteins, mustard proteins, porcine haemoglobins, N,N-dimethylated caseins, bovine beta-lactoglobulins, bovine serum albumins, swine serum albumins, lysozymes, or proteins modified with a chromophore.

10. The method of claim 8 , wherein the sample is a biological sample.

11. The method of claim 10 , wherein the biological sample is an enzyme preparation comprised of a fermentation broth, a feed or in an extract of the fermentation broth or the feed.

12. The method of claim 8 , wherein determining if there is a detectable change in the detectable property of the medium comprises one or more of a quantitative assay, a semi-quantitative assay or a qualitative assay.

Assignees

• Yu Shukun
• Dupont Nutrition Biosci Aps

Inventors

• Yu Shukun

Classifications

• G01N33/5758

  involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites · CPC title

• G01N33/56966

  Animal cells · CPC title

• G01N33/491

  by separating the blood components (G01N15/05 takes precedence) · CPC title

• C12Q1/68

  involving nucleic acids · CPC title

• C12Q1/42

  involving phosphatase · CPC title