Preparation of oligomeric xylose and alpha-glucuronylic enzyme used therefor

A technology of glucuronidase and xylooligosaccharides, which is applied in the field of preparation of xylooligosaccharides, can solve problems such as inability to combine and decompose, affect the quality of xylooligosaccharides, and affect the efficiency of enzymatic hydrolysis

Inactive Publication Date: 2005-02-23
JIANGNAN UNIV
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  • Summary
  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

However, about 94% of the hemicellulose in agricultural and forestry wastes such as corncobs, bagasse and straw is arabinose glucuronoxylan, which is a chain linked by β-1,4 glycosidic bonds. Xylan backbone with side branches composed of 1,3-arabinose or 1,2-glucuronic acid. When a single xylanase is used to hydrolyze this type of xylan to produce xylooligosaccharides, α-glucose Aldehydic side branches create a spatial barrier to the action of xylanase, making xylanase unable to bind and decompose xylan adjacent to these side branches, which not only affects the efficiency of enzymatic hydrolysis, but also affects the quality of xylooligosaccharides

Method used

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  • Preparation of oligomeric xylose and alpha-glucuronylic enzyme used therefor
  • Preparation of oligomeric xylose and alpha-glucuronylic enzyme used therefor
  • Preparation of oligomeric xylose and alpha-glucuronylic enzyme used therefor

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Embodiment 1

[0030] Embodiment 1: refer to Figure 6And other accompanying drawings, the concrete operation of the present invention is as follows:

[0031] (1) Extract corn cob xylan with alkali.

[0032] Take the air-dried corn cob fiber with a particle diameter of about 3mm, first soak it in clean water and then centrifuge to remove the water, add sodium hydroxide solution according to the solid-liquid ratio of 7-10:1, and the final concentration is 1%-3%, at room temperature or high temperature Then, the filtrate was collected and neutralized to slightly acidic with acid, and then precipitated with three times the volume of ethanol to obtain xylan.

[0033] (2) Preparation of enzymes

[0034] Two pairs of primers were designed according to the sequences of xynB and aguA in Genbank, and the genomic DNA of T. maritima was used as a template for PCR amplification. Tm-xynB-N-terminal primer is 5'-CCGTTCCATGGACTACAGGATGTGC-3', Tm-xynB-C-terminal primer is 5'-CCGCTCGAGCGGATATATCTTTCTTCCCT...

Embodiment 2

[0043] Embodiment 2: substantially the same as Example 1, the difference is that xylan is treated with 25mmol / L pH6.3Na 2 HPO 4 -NaH 2 PO 4 The buffer solution is formulated at a concentration of 20%, and the enzyme dosage is: 200 U / g of thermostable xylanase, 15 U / g of thermostable α-glucuronidase, hydrolysis temperature of 80° C., hydrolysis for 12 hours.

Embodiment 3

[0044] Embodiment 3: substantially identical with embodiment 1, difference is that xylan is used 40mmol / L pH6.0Na 2 HPO 4 -NaH 2 PO 4 The buffer solution is prepared at a concentration of 15%, and the enzyme dosage is: 300 U / g of thermostable xylanase, 30 U / g of thermostable α-glucuronidase, hydrolysis temperature of 75° C., hydrolysis for 5 hours.

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Abstract

A method for preparing oligoxylose and alpha-glucose aldehydic acid enzyme is disclosed. It includes: saccharogenic decomposing xylan by combination of xylanase and alpha-glucose aldehydic acid enzyme, expanding and obtainingalpha-glucose aldehydic acid enzyme gene from donor bacteria by PCR, inserting carrier, constructed recombinant plasmid converting into host cell and expressing it, centrifugal obtaining supernatant fluid(enzyme liquid) by centrifugal collecting cell, breaking cell and heat treating. It achieves high zymoprotein output, activity and purification.

Description

technical field [0001] The invention relates to a method for preparing xylooligosaccharide and a method for preparing α-glucuronidase used therein. Background technique [0002] Xylooligosaccharides, also known as xylooligosaccharides, are composed of 2-7 xyloses connected by β-1,4-glycosidic bonds, mainly xylobiose and xylotriose. Various studies have shown that xylooligosaccharides, especially xylobiose, have significant bifidobacteria proliferation ability, non-digestibility, no dental caries and promote the absorption of calcium by the human body. Therefore, xylooligosaccharides are used as A new type of functional oligosaccharide has become one of the hot spots in the research and development of the food industry. Our country is a large agricultural country, which produces 570 million tons of various crop straws every year. If it is only burned for fertilizer, it will waste resources and cause environmental pollution. Therefore, using these biological wastes to prepar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/00C12P19/12
Inventor 薛业敏邵蔚蓝毛忠贵
Owner JIANGNAN UNIV
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