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Method for preparing xyloglucan-oligosaccharides

A technology of xyloglucan oligosaccharide and xyloglucan, which is applied in the field of preparing oligosaccharides and can solve the problems of increasing calorie content and the like

Pending Publication Date: 2020-10-27
TECH UNIV MUNCHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, they can be metabolized by the body during intestinal passage, thus greatly increasing the calorie content of such foods

Method used

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  • Method for preparing xyloglucan-oligosaccharides
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  • Method for preparing xyloglucan-oligosaccharides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0210] Example 1: Cloning of SEQ ID NO:11-14 encoding endoglucanase and expression of SEQ ID NO:1-4

[0211] All chemicals used as buffers and substrates were commercial products of at least reagent grade.

[0212] Cloning of SEQ ID NO: 2: A novel cellulose-degrading bacterial strain was isolated from a 20-liter fermenter operated at 55°C with cow dung and fed with corn silage, and named Herivorax saccincola DSM101079. Herbivorax saccincola was classified as a new family within Ruminococcaceae (Koeck et al. 2016). To carry out genome sequencing to strain Herbivorax saccincola DSM101079, a total of 4 μg of genomic DNA was used to construct an 8-k end paired (mate-pair) sequencing library (Nextera Mate Pair Sample Preparation Kit, Illumina Inc.) End-of-Protocol to sequence it. The analysis and interpretation of the Herbivorax saccincola DSM101079 genome sequence in GenDB, and through the Carbohydrate-active-enzyme database dbCAN (Yin et al., 2012) revealed more than 100 genes,...

Embodiment 2

[0221] Example 2: Characterization of SEQ ID NO: 1-4

[0222]Enzyme specific activities were determined by using model substrates such as barley-β-glucan (Megazyme). Endoglucanase activity is defined as the production of reducing sugars from model substrates. One unit of enzyme activity is defined as the amount of micromoles of reducing sugar produced per mg of enzyme in one minute at 60°C. To determine the zymogram and optimum temperature and pH, endoglucanase (50 ng / reaction) was incubated at a temperature range of 50 to 80 °C in the presence of 1% (wt / vol) barley-β-glucan Incubate for 30 minutes under conditions, the barley-β-glucan was dissolved in citrate buffer with different pH (range 4.0-8.0) (solution A: 0.2M citric acid, 0.1M NaCl; solution B: 0.4M Na 2 HPO 4 , 0.1M NaCl). Measurement of reducing sugars by 3,5-dinitrosalicylic acid (DNSA) method: Mix 50 μl of sample with 75 μl of DNSA solution (10 g / L DNSA, 200 g / L K + -Na + - Tartrate, 10g / L NaOH, 0.5g / LNa 2...

Embodiment 3

[0225] Example 3: Hydrolysis of TKP

[0226] Tamarind kernel powder (TKP) and deoiled tamarind kernel powder (dTKP) were purchased from Tamarind Magic, Hyderabad, Phase-IV, 3rd Gate, IDA, Cherlapally, Hyderabad-500051, Telangana, India. Both TKP and dTKP consist of about 65% (wt / wt) xyloglucan (https: / / www.altrafine.com / tamarind-kernel-powder / ).

[0227] For XGOS release, 8 g of TKP or dTKP at 20% (wt / vol) was dissolved in 40 ml of demineralized water at 60°C. Enzymes were added at desired concentrations of 3%, 0.65%, 0.05%, 0.03% or 0.006% relative to substrate loading (wt / wt TKP). The reaction mixture was incubated on a rotary shaker at 125 rpm and 60°C. The hydrolysis process was monitored by taking samples at different time points. Samples were centrifuged and supernatants were boiled at 95°C for 5 minutes to denature residual proteins. For analysis, supernatant samples were taken, diluted tenfold with double deionized water and measured by HPAEC-PAD.

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Abstract

The present invention relates to a method for preparing oligosaccharides which can be used among others as food additives to reduce calorie content, to sweeten food products, to increase the fiber content of food products, to improve the texture of food products and to stimulate the gut microbiome bacteria. Furthermore they can be applied in the fields of animal feed, or other applications. More particularly, this invention is directed to a high temperature hydrolysis of xyloglucan polysaccharide to defined xyloglucan oligosaccharides. The invention further relates to oligosaccharide hydrolysates produced with the method of the invention and to the use of said oligosaccharide hydrolysates in human and / or animal nutrition, as prebiotic or other uses. Further provided are novel endoglucanases for use in the method of the invention as well as in other applications.

Description

technical field [0001] The present invention relates to a process for the preparation of oligosaccharides which can be used inter alia as food additives to reduce the calorie content of food products, to sweeten food products, to increase the fiber content of food products, to improve the texture of food products and to stimulate intestinal tract Tract microbiome bacteria. In addition, they can also be used in animal feed or other applications. More specifically, the present invention relates to the pyrohydrolysis of xyloglucan polysaccharides to defined xyloglucanoligosaccharides. The present invention also relates to oligosaccharide hydrolysates produced by the method of the invention, and to the use of said oligosaccharide hydrolysates as prebiotics or other uses in human and / or animal nutrition. Also provided are novel endoglucanases for use in the methods of the invention, among other applications. Background technique [0002] In general, fats, oils, starches, carbo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/716C12P19/14C12N9/42A23L29/206
CPCA61K31/702C12P19/14C12N9/2402C12N9/2437C12N9/2451C12Y302/01004A23L29/206C12Y302/01151
Inventor S·格劳布纳V·兹韦尔洛夫W·施瓦茨P·科恩贝格尔B·安德烈森
Owner TECH UNIV MUNCHEN
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