Method for producing xylobiose, and dedicated immobilized enzyme

A technology for immobilizing enzymes and xylanases, applied in directions such as hydrolase, bacteria, etc., which are immobilized on/in organic carriers, can solve the problems of low proportion of xylobiose, and achieve easy control of product composition, The effect of high recovery of enzyme activity and high stability

Inactive Publication Date: 2010-05-12
CHINA AGRI UNIV
View PDF11 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods can prepare xylooligosaccharides, but the proportion of xylobiose is low

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
  • Method for producing xylobiose, and dedicated immobilized enzyme
  • Method for producing xylobiose, and dedicated immobilized enzyme
  • Method for producing xylobiose, and dedicated immobilized enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, preparation immobilized xylanase

[0042] 1. Preparation of Eupergit C 250L-Ni immobilized XynB

[0043] 1. Preparation of XynB enzyme solution

[0044] The crude enzyme solution was obtained according to the method described in the first paragraph of Example 2, Example 3, Example 4, and Example 5 of Chinese Patent ZL 02156022.6. The specific method is as follows:

[0045] (1) Amplification of xylanase gene xynB

[0046] The primers used are TM-XynB-Nco I-FWD and TM-XynB-Hind-His-REV, and their sequences are as follows:

[0047] TM-XynB-Nco I-FWD: 5'-CCATGGAAA TATTACCTTC TGTGTGAT (SEQ ID NO: 2)

[0048] TM-XynB-Hind-His-REV: 5'-AAGCTTT CTTTCTTCTA TCTTTTTCTC CA (SEQ ID NO: 3)

[0049] According to the complete sequence of the xynB gene of Thermotoga maritima MSB8 DSM3109 preserved in the German Microorganism Collection (DSM), the above-mentioned pair of primers were designed, and the pET28a (+) (Novagen, USA) plasmid capable of inserting into the prime...

Embodiment 2

[0087] Embodiment 2, the preparation of steam explosion liquid or high temperature cooking liquid

[0088] 1. Preparation of corn cob steam explosion liquid or high temperature cooking liquid

[0089] 1. Preparation of steam explosion liquid

[0090] Weigh 100g corn cob (containing 32.4% xylan), add 800mL of distilled water according to the solid-to-liquid ratio of 1:8, soak at room temperature for 12h, put it into a high-pressure tank, cover and seal, heat, and keep the temperature at 165-170°C for 30min Immediately after opening the ball valve, the material is released into the collection tank. When the temperature of the collected material drops to 60-70°C, it is sucked and filtered, and the filtrate is the steam explosion liquid. After cooling to room temperature, determine its total sugar and reducing sugar content. Among them, the content of reducing sugar was determined by Somogyi method with D-xylose as the standard. The total sugar was determined by the Orcinol-HC...

Embodiment 3

[0095] Embodiment 3, using the immobilized xylanase of embodiment 1 to continuously hydrolyze the steam explosion liquid or high-temperature cooking liquid of embodiment 2

[0096] 1. Hydrolysis of Eupergit C 250L-Ni immobilized XynB column treated with 1M phosphate buffer Example 2 Corn cob steam explosion liquid or high temperature cooking liquid

[0097] The experiment set up 3 repetitions. Each repetition added 40g wet state (prepared by 10g Eupergit C 250L) 1M phosphate buffered Eupergit C 250L-Ni immobilized XynB to a stainless steel jacketed chromatography column with an internal diameter of 15mm, the actual height of the enzyme bed was 33cm, and the connection constant flow pump and fraction collector ( Figure 5 ), equilibrate with 50mM, pH 6.2 citric acid buffer for 30min. The substrate is corncob steam-popping liquid or high-temperature cooking liquid, wherein the total sugar content of corncob steam-popping liquid is 22.0mg / mL, and the reducing sugar content is 4....

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

PropertyMeasurementUnit
eccentricityaaaaaaaaaa
pore sizeaaaaaaaaaa
melting pointaaaaaaaaaa
Login to view more

Abstract

This invention discloses a method for preparing xylobiose and its specific immobilization enzyme. The method comprises: (1) immobilizing recombined xylanase XynB using metal-chelating epoxy carrier Eupergit C 250 L to obtain immobilization enzyme; (2) suspending the obtained immobilization enzyme in 0.5-1.25 M phosphate buffer with a pH of 5.5-8.0, and reacting at 20-35 deg.C for 12-60 h to obtained immobilization xylanase. The recombinant xylanase XynB is prepared by expressing xylanase gene (Sequence ID 1) in Escherichia coli. The obtained xylanase has such advantages as high recovery rate;high stability and good operation performance after loaded into the column, and can be used for continuous production. The method for preparing xylobiose by using xylanase is simple, and can save xylanase usage, realize separation of enzyme and hydrolyte, largely lower production cost and improve production efficiency, thus is suitable for mass production.

Description

technical field [0001] The invention relates to a method for producing xylobiose and its special immobilized enzyme. Background technique [0002] Xylobiose (Xylobiose) has unique physiological functions and good process performance. The sweetness is about 40% of sucrose, and the sweetness is refreshing and soft. It can be used in many fields such as food, medicine and health products. Active ingredients, which are also qualitative and quantitative components of xylooligosaccharides, are a hotspot in the research of functional oligosaccharides. Its main functional characteristics are as follows: 1) Significant bifidobacteria proliferation ability, which can rapidly proliferate beneficial bacteria such as bifidobacteria, thereby inhibiting the growth of harmful bacteria in the intestinal tract. 2) Indigestibility: Xylobiose is very stable in the human digestive system and cannot be decomposed by saliva, pancreatic juice, small intestinal villous gland homogenate, etc. It ca...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/02C12N9/24C12N1/21C12P19/12
Inventor 江正强李里特李道义朱运平
Owner CHINA AGRI UNIV
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