Methods for detecting and measuring polysaccharide-hydrolyzing enzymes

Inactive Publication Date: 2010-09-23
SYNGENTA PARTICIPATIONS AG
View PDF6 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0008]Compositions and methods are provided for measuring a polysaccharide-hydrolyzing enzyme in a sample, particularly in a recombinant plant sample. The methods involve measuring changes in chemical and physical properties of the sample, such as viscosity, to detect and/or measure a polysaccharide-hydrolyzing enzyme in the sample. The viscosity change

Problems solved by technology

Current methods for detecting and measuring transgenic plant material containing a polysaccharide-hydrolyzing enzyme are time consuming in that they require milling the plant material, extracting the polysaccharide-hydrolyzing enzyme from a plant sample, adding exogenous polysaccha

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Methods for detecting and measuring polysaccharide-hydrolyzing enzymes
  • Methods for detecting and measuring polysaccharide-hydrolyzing enzymes
  • Methods for detecting and measuring polysaccharide-hydrolyzing enzymes

Examples

Experimental program
Comparison scheme
Effect test

example 1

Modified Fast Assay for Corn Amylase

I. Enzyme Calibration Curve Construction Using % Admix.

[0071]1) Creation of Calibration Samples

[0072]Corn Event 3272 (as described in U.S. 2006 / 0230473 comprising the thermotolerant alpha-amylase 797GL3) and commodity corn (negative background) (i.e. Yellow dent) was ground separately (i.e., dry milled) to flour in a Perten LM3600 Disc Mill (setting 0) (Perten Instruments AB; Huddinge, Sweden). Moisture content was measured using a Mettler Halogen Moisture Analyser to calculate total dry weight (data not shown). Admixtures of ground Event 3272 seed was mixed into ground commodity corn to make admixes of 40%, 50%, 60%, 70%, 80%, 90% and 100% using a calculated total dry weight of 500 g.

[0073]2) Viscosity Measurement

[0074]Approximately 9-20 grams of each admixture was weighed separately into viscometer vessels. Water was next added to each tube to create a slurry containing about 28% dry solids. Corn slurry has very good buffering capacity, so it is...

example 2

Modified Fast Assay for Corn Amylase Using a Standard Coffee Maker

[0091]1) Creation of Calibration Samples

[0092]Calibration samples were created essentially as described in (Example 1) above. Corn Event 3272 (comprising the thermotolerant alpha-amylase 797GL3) and commodity corn (negative background) (i.e. Yellow dent) were ground separately (i.e., dry milled) to flour in a Perten LM3600 Disc Mill (setting 0) (Perten Instruments AB; Huddinge, Sweden). Moisture content was measured using a Mettler Halogen Moisture Analyser to calculate total dry weight (data not shown). Admixtures of ground Event 3272 seed was mixed into ground commodity corn to make admixes of 40%, 50%, 60%, 70%, 80%, 90% and 100% using a calculated total dry weight of 500 g.

[0093]2) Generation of a Calibration Curve

[0094]Approximately 15 grams form each admix was added separately to a coffee filter fitting a standard 4 cup coffee maker such as a Mr. Coffee® 4-Cup Dispenser (Sunbeam Products, Inc.) (or any non-progr...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

Methods are disclosed for detecting and measuring polysaccharide-hydrolyzing enzyme activity or concentration by partial hydrolysis using a pre-determined, yet short, incubation time and a pre-determined temperature. The resulting reaction mixture has unique chemical (i.e., reaction products) and physical (i.e., viscosity) properties that can be used to detect or measure the polysaccharide-hydrolyzing enzyme activity or concentration.

Description

RELATED APPLICATIONS[0001]The following application claims priority to U.S. Provisional Application No. 61 / 161,182; filed Mar. 18, 2009FIELD OF THE INVENTION[0002]The invention relates generally to methods of detecting and measuring a polysaccharide-hydrolyzing enzyme and more particularly to methods of detecting and measuring the polysaccharide-hydrolyzing enzyme by observing changes in chemical and physical properties of a polysaccharide-containing sample at a pre-determined temperature over a pre-determined time period.BACKGROUND OF THE INVENTION[0003]Polysaccharides, such as starches, are complex carbohydrates composed of monosaccharides joined via glycosidic bonds. They are typically amorphous and insoluble in water. Examples of polysaccharides include storage polysaccharides such as starch and glycogen, and structural polysaccharides such as cellulose and chitin.[0004]Starch is a major carbohydrate reserve in plant tubers and seed endosperm. The largest source of starch is cor...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/40C12Q1/34
CPCC12Q1/34C12Q1/40G01N2333/944G01N2333/928G01N2333/942G01N2333/924
Inventor MOSER, TRICIABASU, SHIB SANKARPRATA, ROGERIO T.N.MOON, HANGSIKZHONG, XUEJUNKIM, MYOUNG
Owner SYNGENTA PARTICIPATIONS AG
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap