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High-temperature-resistant alkaline xylanase

A technology of xylanase and high temperature resistance, applied in the directions of glycosylase, enzyme, hydrolase, etc., can solve the problem that it is difficult to achieve the research and development goals and requirements of high temperature resistance, the range of alkali resistance and high temperature resistance is not wide, and the results are not satisfactory. and other problems, to achieve the effect of enhancing the body's functionality, improving the anti-stress ability, adjusting and repairing the body's function

Active Publication Date: 2014-06-18
HUNAN HONGYING BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] From this point of view, many international and domestic experts, scholars and scientific research staff have spared no expense in money, manpower and time to increase the research and development of high temperature and alkali resistant xylanase, aiming to apply biotechnology to papermaking, food , Feed and other fields to a higher, newer, stronger new era, whether it is the acquisition of new strains, or the use of genetic recombination technology, although such a large amount of financial resources and energy have been spent, the results are not satisfactory , it is difficult to achieve the research and development goals and requirements of high temperature resistance and alkali resistance xylanase, either temperature resistance is not alkali resistance; or alkali resistance is not temperature resistance; or the range of alkali resistance and high temperature resistance is not wide; And have cellulase activity; or the substrate is not suitable, etc., to prepare a kind of stronger alkali resistance, higher temperature resistance, wide range of alkali resistance and high temperature resistance, no cellulase activity and suitable for papermaking, food and feed, etc. The high temperature resistant alkaline xylanase in the field is still the relentless pursuit and research and development direction of those skilled in the art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Determination of enzyme activity of high temperature resistant alkaline xylanase (3,5-dinitrosalicylic acid colorimetric method, referred to as DNS method)

[0046] 1. Reagents and solutions

[0047] 1.1 Except for special requirements, the water used is third-grade water in accordance with the provisions of GB / 6682-1992, and the chemicals are all analytically pure unless otherwise specified.

[0048] 1.2DNS Reagent

[0049] Weigh 10g of 3,5-dinitrosalicylic acid and add it into 500ml of water, add 16g of sodium hydroxide in several times, stir to dissolve (temperature <45°C), then add 300g of potassium sodium tartrate in several times, stir until completely dissolved, After cooling, dilute to 1000ml with water. Store the brown bottle in a dark place at room temperature and use it after one week (if there is precipitation, use it after filtration).

[0050] 1.30.1mol / L, pH5.0 acetic acid-sodium acetate buffer

[0051] Take 1.8ml of glacial acetic acid, add appropria...

Embodiment 2

[0079] A preparation method for high temperature resistant alkaline xylanase, comprising the steps of:

[0080] (1) Activation of strains

[0081] Inoculate the slant strain of the well-preserved thermophilic Bacillus 701 on the slant medium, culture at 45°C for 36 hours to activate the strain, and activate it 3 times;

[0082] The slant medium consists of: 10g of beef extract, 12g of sodium chloride, 20g of peptone, 5g of glucose, 20g of agar, 10g of Chinese herbal medicine powder, 1000mL of distilled water, pH 10, and sterilized at 121°C for 20 minutes;

[0083] The preparation method of described Chinese herbal medicine powder is as follows:

[0084] In parts by weight, weigh 30 parts of Radix Astragali, 18 parts of Codonopsis Radix, 15 parts of Bupleurum Radix, and 15 parts of Scutellaria Baicalensis; respectively crush the above-mentioned Chinese herbal medicines to a particle size below 2 mm, then uniformly mix in a container and add 6 times the weight of Water, keep t...

Embodiment 3

[0110] The thermostability analysis of embodiment 3 alkaline xylanase

[0111] The thermal stability of the enzyme was analyzed, and the crude enzyme solution of Example 2 was placed at 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, and 70°C, and samples were taken every 10 minutes to determine the enzyme activity. . At 40°C, 45°C, 50°C, and 55°C in the crude enzyme solution, the enzyme activity did not decrease for 60 minutes. At 60°C and 65°C, the original enzyme activity is reduced to 95% in 30 minutes, and to 85% in 60 minutes. At 70°C, it will drop to 85% of the original enzyme activity in 30 minutes and 80% in 60 minutes. At 75°C, the enzyme activity is reduced to 80% of the original enzyme activity in 30 minutes, and to 70% in 60 minutes. Compared with the prior art, the same enzyme activity is achieved, and the temperature tolerance is higher.

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Abstract

The invention discloses high-temperature-resistant alkaline xylanase and belongs to the technical field of preparation of enzymic preparations. The high-temperature-resistant alkaline xylanase is prepared by taking bacillus thermophilus 701 for producing the high-temperature-resistant alkaline xylanase as an original strain and performing liquid submerged fermentation. The specific activity of the high-temperature-resistant alkaline xylanase is 350-420IU / ml, the applicable temperature range is 40-75 DEG C, the optimal reaction temperature is 65 DEG C, the enzyme activity at the temperature of 75 DEG C is completely stable, the applicable reaction pH value ranges from 5.0 to 11.0, the enzyme activity at the pH value of 11.0 is completely stable, and the optimal reaction pH value is 10.0. Compared with the conventional high-temperature-resistant alkaline xylanase, the high-temperature-resistant alkaline xylanase in the invention is high in enzyme activity, high in temperature resistance, high in alkali resistance and wide in enzyme action optimal pH value range and is particularly suitable for industrial requirements in the fields of papermaking, foods and feed.

Description

technical field [0001] The invention belongs to the technical field of enzyme preparation, in particular to a high-temperature resistant alkaline xylanase. Background technique [0002] Cellulose, hemicellulose and lignin are the main components of plant hemicellulose. They work together to form the supporting framework of plant cell walls. Among the three, hemicellulose accounts for more than 35% of the dry weight of plants. Hemicellulose It is also a general term for a variety of complex glycans, and its main component is xylan, which is a complex poly-five-carbon sugar. The main chain of xylan is connected by multiple xylopyranosyl groups through β-1-4 glycosidic bonds, and different substituents are connected to the side chains, such as acetyl group, glucuronyl group, L-arabinosyl group etc. (Collins et al., 2005). Therefore, the complete degradation of xylan requires the participation of various enzymes. Xylans are generally divided into hardwood xylans and softwood ...

Claims

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Application Information

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IPC IPC(8): C12N9/42D21C9/10C12R1/01
CPCC12N9/2482C12Y302/01008D21C9/10
Inventor 李洪兵朱永明孙明芳刘波易继云周俊
Owner HUNAN HONGYING BIOTECHNOLOGY CO LTD
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