A kind of preparation method of high temperature resistant alkaline xylanase

A technology of xylanase and high temperature resistance, which is applied in microorganism-based methods, biochemical equipment and methods, glycosylase, etc. Broad, unsatisfactory results, etc., to achieve the effect of enhancing the body's functionality, improving the ability to resist stress, adjusting and repairing the body's function

Active Publication Date: 2015-09-16
南宁雄晋生物科技有限公司
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  • Summary
  • 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

[0051] The thermophilic bacillus 701 producing high-temperature-resistant alkaline xylanase provided by the present invention is a strain of thermophilic bacillus HYX0021 isolated from soil samples collected from Xueli Paper Mill in Hunan Province. After repeated ultraviolet mutagenesis and nitroso Obtained by guanidine mutagenesis screening, it is characterized by high temperature resistance and strong alkali resistance.

[0052] The strain producing thermostable alkaline xylanase provided by the present invention is specifically thermophilic Bacillus 701. The strain was deposited on October 12, 2013 at the China Center for Type Culture Collection (CCTCC for short, address: School of Life Sciences, Wuhan University, Luojia Mountain, Wuchang District, Wuhan City, Hubei Province, Zip Code: 430072), and the preservation number is CCTCC M2013537.

[0053] The slant medium consists of: beef extract 3-10g, sodium chloride 5-12g, peptone 10-20g, glucose 2-5g, agar 15-20g, Chinese he...

Embodiment 2

[0066] A preparation method for high temperature resistant alkaline xylanase, comprising the steps of:

[0067] (1) Activation of strains

[0068] Inoculate the slant strain of the well-preserved thermophilic Bacillus 701 on the slant medium, culture at 37°C for 24 hours to activate the strain, and activate twice in this way;

[0069] The slant medium consists of: 3g of beef extract, 5g of sodium chloride, 10g of peptone, 2g of glucose, 15g of agar, 5g of Chinese herbal medicine powder, 1000mL of distilled water, pH 5, and sterilized at 121°C for 20 minutes;

[0070] The preparation method of described Chinese herbal medicine powder is as follows:

[0071] In parts by weight, weigh 20 parts of Astragalus membranaceus, 10 parts of Codonopsis pilosula, 10 parts of Bupleurum radix, and 10 parts of Scutellaria baicalensis, respectively crush the above-mentioned Chinese herbal medicines to a particle size below 2 mm, then uniformly mix them in a container and add 3 times the weigh...

Embodiment 3

[0099] A preparation method for high temperature resistant alkaline xylanase, comprising the steps of:

[0100] (1) Activation of strains

[0101] Inoculate the slant strain of the well-preserved thermophilic Bacillus 701 on the slant medium, culture at 41°C for 30 hours to activate the strain, and activate it 3 times;

[0102] The slant medium consists of: 6g of beef extract, 8g of sodium chloride, 15g of peptone, 5g of glucose, 20g of agar, 8g of Chinese herbal medicine powder, 1000mL of distilled water, pH 8, and sterilized at 121°C for 20 minutes;

[0103] The preparation method of described Chinese herbal medicine powder is as follows:

[0104] In parts by weight, weigh 25 parts of Radix Astragali, 15 parts of Codonopsis pilosula, 12 parts of Bupleurum, and 12 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 5 times the weight of Water, keep the temp...

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Abstract

The invention discloses a method for preparing high-temperature-resistant alkaline xylanase, belonging to the technical field of preparation of enzyme preparations. Bacillus thermophilus 701 capable of producing high-temperature alkaline xylanase is taken as a starting strain, and high-temperature-resistant alkaline xylanase, which has strong high-temperature resistance and alkali resistance, wide acting conditions, no cellulase activity and is suitable for the fields such as papermaking, food and feed, is prepared by optimizing the culture medium composition and fermentation process in a way of liquid submerged fermentation. The specific activity of the enzyme is 350-420IU / ml; the optimal reaction temperature is 65 DEG C, the application temperature range is 40-75 DEG C; the optimal reaction pH value is 10.0 and the pH range applicable to reaction is 5.0 to 11.0.

Description

technical field [0001] The invention belongs to the technical field of enzyme preparation, in particular to a preparation method of 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 hardwoo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42D21C9/10C12R1/07
CPCC12N9/2482C12Y302/01008D21C9/10
Inventor 李洪兵李海清胡永明刘文明易继云雷敏
Owner 南宁雄晋生物科技有限公司
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