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The preparation method of xylooligosaccharide

A technology for xylo-oligosaccharide and xylan, which is applied in the field of preparation of xylo-oligosaccharide, can solve the problems of low enzymatic hydrolysis efficiency, high process cost, difficult to be enzymatically hydrolyzed, etc., and achieves improved enzymatic hydrolysis efficiency, low price, and reduced process cost effect

Active Publication Date: 2021-01-22
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the natural xylan molecule is a hybrid polysaccharide, the cross-linking between the main chain and the side chain molecules causes xylan to form a complex networked substrate structure that is difficult to be enzymatically hydrolyzed. The process has problems such as low enzymatic hydrolysis efficiency and high process cost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1) Preparation of high-temperature-resistant endoxylanase: Heat-shock transformation of recombinant plasmid pET-28b-CoXynA constructed with high-temperature-resistant endoxylanase gene into Escherichia coli Transetta (DE3) host to construct and express high-temperature-resistant Genetically engineered strains of endoxylanase. Cultivate the above-mentioned genetically engineered bacteria at 37°C, when the OD of the bacterial solution 600 When the value reaches 0.8, induce with isopropylthiogalactopyranoside (IPTG) for 6 hours at 37°C, and then break the bacteria by high-pressure homogenization, and then pass the supernatant of the bacteria liquid through affinity chromatography. A high-temperature-resistant endoxylanase solution (5.1 mg / mL) was prepared after a column (nickel column), anion exchange column and molecular sieve (Superdex75). The number of thermostable endoxylanase in NCBI database is ADQ03732.

[0026] 2) Preparation of bifunctional xylan hydrolase: The ...

Embodiment 2

[0031]Xylooligosaccharides were prepared using the same strategy as in Example 1, except that different pretreatment substrates were used: bagasse was crushed, dried, passed through a 60-mesh sieve, and weighed. The product was analyzed by HPLC and contained 279 mg of xylooligosaccharides, including 57 mg of xylobiose, 159 mg of xylotriose and 63 mg of xylotetraose.

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Abstract

The invention discloses a method for preparing xylo-oligosaccharide. The method includes that natural xylan is used as an enzymatic hydrolysis substrate, and enzymatic hydrolysis is sequentially carried out by high-temperature-resistant xylan endonuclease and dual-function xylanse. The high-temperature-resistant xylan endonuclease at least comprises amino acid fragments V-F-E-F-E-D-R-R-T-D-L-L-E-P-A-E, and the dual-function xylanse at least comprises amino acid fragments E-N-Q-N-D-I-P, G-G-G, F-S-A, D-D-P-V-E-R, D-D-S and R-H-G. The method has the advantages that the substrate does not need tobe chemically pretreated, and accordingly the method is high in enzymatic hydrolysis efficiency and low in process cost.

Description

technical field [0001] The invention relates to a preparation method of xylooligosaccharide. More specifically, the present invention relates to a method for the preparation of xylooligosaccharides without chemical pretreatment of the substrate. Background technique [0002] Xylooligosaccharides are widely used in the health food industry as prebiotics, and can also be used in foods as humectants or low-calorie additives. The by-products of agricultural product processing are rich in xylan resources. Xylooligosaccharide is a mixture of xylooligosaccharides with xylobiose, xylotriose and xylotetraose as the main components obtained by hydrolyzing β-1,4 xylosidic bonds with xylan as a substrate. . Corncob, bagasse, and rice husk all have high xylan content, which are ideal natural raw materials for the preparation of xylooligosaccharides. However, since the natural xylan molecule is a hybrid polysaccharide, the cross-linking between the main chain and the side chain molecu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/00C12P19/12C12P19/14
Inventor 辛凤姣曹浩温博婷王凤忠孙丽超谢莹莹谷天一
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI