Method for producing low polyxylose with dual-aqueous phase hydrolyzation system

A technology of xylooligosaccharide and hydrolysis system, which is applied in the direction of fermentation, etc., can solve the problems of difficult scale-up, low recycling rate of xylanase, and many process steps, so as to achieve high utilization rate, simplify the enzyme hydrolysis process, and simplify the process route effect

Inactive Publication Date: 2008-10-29
QINGHAI WEIDE SPECIAL SUGAR
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

Although the enzymatic hydrolysis method has obvious advantages, there are also problems such as low recycling rate of xylanase, many process steps, and difficulty in scaling up the bioprocessing process.

Method used

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  • Method for producing low polyxylose with dual-aqueous phase hydrolyzation system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: the preparation of xylanase

[0034] The xylanase used in the present invention is derived from recombinant Pichia pastoris NL-OU (Ouyang Jia, Wang Xiangming, Zhang Qing, Yan Ming, Xu Lin. Trichoderma reesei xylanase XYN II gene in Pichia pastoris Secreted expression. Forest Products Chemistry and Industry, 2007, 27(5): 83-88). The preparation method is as follows: After the recombinant Pichia pastoris NL-OU is activated on a plate, inoculate a single colony in a 50mL shake flask medium, and culture it at 30°C and 150r / min for 48 hours until the OD 600 Reach 2-6, obtain seed solution. Inoculate the seed solution with 10% (v / v) into sterilized glycerol medium, and ferment for about 20 hours at 28°C and 600r / min until the dissolved oxygen rises by more than 50%. Glycerol solution was fed into the tank at a flow rate of 20 mL / (L·h) for 12 hours, then stopped for 3 hours, the pH value was controlled at about 5.0, and the temperature was maintained at 28°C. Th...

Embodiment 2

[0035] Embodiment 2: the preparation of xylan

[0036] The corncobs are dried, crushed and sieved to obtain raw materials with particles of about 1 cm. At 25°C, sodium hydroxide and deionized water were added and stirred while adding. Then, cook at 90° C. for 3 hours. Filter to remove solids and collect cooking liquid. The cooking liquid is neutralized to pH5.0 with strong acid. The neutralized cooking liquid is desalinated by an ultrafiltration membrane with a molecular weight cut-off of 6000, and more than 90% of inorganic salts and low-molecular compounds are removed to obtain concentrated crude xylan liquid.

Embodiment 3

[0037] Example 3: Preparation of xylo-oligosaccharides by aqueous two-phase enzymatic hydrolysis

[0038] In the hydrolysis tank of the two-phase hydrolysis reactor, add concentrated crude xylan solution 200mL (xylan concentration is 20g / L), add 150g polyethylene glycol 2000 and 100g ammonium sulfate, add deionized water to total The system weight is 1000g. In the polyethylene glycol / ammonium sulfate system, the concentration of polyethylene glycol 2000 is 15% (w / w), and the concentration of ammonium sulfate is 10% (w / w). After mixing by mechanical stirring (150r / min), add 200IU xylanase crude enzyme solution for enzymatic hydrolysis. Under the conditions of pH 5.0, temperature 50° C., and stirring speed 150 r / min, the enzymatic hydrolysis reaction was carried out for 4 hours. Then, transfer the enzyme hydrolyzate to the distribution tank of the two-phase hydrolysis reactor, and let it stand for 60 minutes to complete the phase formation. The lower aqueous solution phase us...

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Abstract

The invention discloses a method for preparing xylooligosaccharides by using a double water-phase hydrolysis system, which comprises the following steps: (1) configuring a double water-phase system containing polyethylene glycol and ammonium sulfate or polyethylene glycol and glucan in a hydrolyzing tank; (2) adding crude xylan solution and xylanase for hydrolyzing; (3) adding the reactive solution to a dispensing groove, standing for separating phases, separating the lower phase by an ultrafiltration membrane, separating with nanofiltration membrane to obtain xylooligosaccharides, recovering the residues obtained by ultrafiltration, mixing with the upper phase and adding into the hydrolyzing tank; and (4) adding ammonium sulfate in the hydrolyzing tank for the original double water-phase system containing polyethylene glycol and ammonium sulfate, returning to step (2), and returning to step (2) for the original double water-phase system containing polyethylene glycol and glucan. The method can recover xylanase by using the double water-phase system, and has the advantages of improved utilization rate of xylanase, simple operation, easy scale up, and low production cost.

Description

technical field [0001] The invention relates to a preparation method of xylo-oligosaccharides, in particular to a method for preparing xylo-oligosaccharides by integrating a xylanase hydrolysis process with a two-water phase separation system. Background technique [0002] Natural cellulose resources are the most abundant renewable organic matter on the earth. The biomass produced through plant photosynthesis in the world is as high as 100 billion tons, of which only 11% are used for crop products, feed, papermaking and building materials, etc., and 89% Natural cellulose resources are degraded by various microorganisms in nature and finally converted into carbon dioxide and water. Although this is an important carbon cycle in nature, it is undoubtedly a huge waste of resources from the perspective of atomic economy. As one of the important components of natural cellulose resources, hemicellulose is the most abundant and cheapest renewable resource besides cellulose. Using ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/00C12P19/04C12P19/12
Inventor 李鑫欧阳嘉余世袁勇强宋向阳徐勇陈牧
Owner QINGHAI WEIDE SPECIAL SUGAR
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