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Thermophilic bacillus licheniformis UTM102 for producing phytase and application of thermophilic bacillus licheniformis UTM102

A technology of Bacillus licheniformis and UTM102, which is applied in the field of fermentation to achieve the effects of promoting plant growth, promoting plant growth, and increasing yield

Active Publication Date: 2015-04-29
大地绿源环保科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has not been reported that Bacillus licheniformis with phytase activity and a variety of high-temperature-resistant biomass hydrolytic enzymes is used in the treatment of organic solid waste materials, and the preparation of growth-promoting functional biological fertilizers has not been reported.

Method used

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  • Thermophilic bacillus licheniformis UTM102 for producing phytase and application of thermophilic bacillus licheniformis UTM102
  • Thermophilic bacillus licheniformis UTM102 for producing phytase and application of thermophilic bacillus licheniformis UTM102
  • Thermophilic bacillus licheniformis UTM102 for producing phytase and application of thermophilic bacillus licheniformis UTM102

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Isolation and identification of Bacillus licheniformis UTM102

[0034] Take 1 gram of hot spring bottom mud sample and place it in a 250ml Erlenmeyer flask containing multiple small glass beads and 50ml sterile water, shake on a constant temperature shaker at 40°C for 1 hour, and let stand for 30 minutes. Aspirate 1ml of the supernatant under aseptic conditions and connect it to a 250ml triangle containing 50ml of sterilized acclimation medium (glucose 10g, beef extract 3g, yeast extract 5g, peptone 10g, sodium chloride 5g, 1000ml distilled water) Incubate in the bottle at 40°C for 3 days (rotational speed 160 rpm). In the same way, draw 1ml of the enrichment solution again and subculture for 3 generations.

[0035] Take 1ml of the bacterial suspension after 3 generations of culture, add it to 9ml of sterile water, and prepare 10 by the 10-fold dilution method. -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 The dilution. 0.1ml of each dilution was spread on 3...

Embodiment 2

[0040] Example 2 Detection of cellulase activity of strain UTM102 and related gene sequence

[0041] Spot the Bacillus UTM102 spot seeding method in the sodium carboxymethyl cellulose medium (sodium carboxymethyl cellulose 5g, KH 2 PO 4 lg, agar 17g, NaNO 3 3g, KCL 0.5g, MgSO 4 0.5g, FeSO 4 0.01g, 1000ml of distilled water, pH 5.5-6.0), incubate at 40°C for 48 hours. Dye with 0.2% Congo red for 30 minutes, wash off the dye solution with distilled water, soak in NaCl with a concentration of 1mol / L for 1 hour, and finally fix the color with 5% acetic acid solution. The formation of a colorless transparent circle around the colony indicates that the bacteria secrete cellulase. Cellulase is a mixture of multiple enzymes, which is composed of cellobiase, β-glucanase and the like.

[0042] According to beta-glucan endonuclease gene design primer betaglu-f: 5'-CGA TGT TGTTCA TGC CGG CT-3' and betaglu-r: 5'-TTG CCA GCG TGT GTG ACAGC-3', with the strain of the present invention UTM102...

Embodiment 3

[0044] Example 3 Detection of xylanase activity of strain UTM102 and its gene sequence

[0045] Accurately weigh the beech wood xylan (Sigma) (accurate to 0.001 g), and use 50mmol / L NaAc-HAc (pH 5.0) buffer to make a 1% beech wood xylan solution (preferably used fresh, glycan It is easy to decompose under acidic conditions); in addition, take 1ml of the supernatant of UTM102 culture (fermentation medium: glucose 5g, peptone 15g, yeast extract 5g, sodium chloride 5g, distilled water 1000ml, pH7.0), use 50mmol / L The NaAc-HAc (pH 5.0) buffer is formulated into an enzyme solution with an appropriate concentration to ensure that the light absorption is between 0.2 and 0.6. Then take 0.9ml of 1% beech xylan solution, add 0.1ml of enzyme solution prepared by UTM102, incubate in a 55℃ water bath for 30 minutes, add 1ml of distilled water and 2ml of DNS reagent. After mixing, boiling in water bath for 5 minutes, cooling to room temperature, adding distilled water to make the volume up to...

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Abstract

The invention provides a thermophilic bacillus licheniformis UTM102 for producing phytase and an application of the thermophilic bacillus licheniformis UTM102. The biological collection number of the thermophilic bacillus licheniformis UTM102 is CGMCC (China General Microbiological Culture Collection Center) No.9508. The thermophilic bacillus licheniformis UTM102 can perform aerobic composting fermentation in organic solid wastes such as sewage sludge, household garbage, animal carcasses, livestock manure, crop straw and the like, can adapt to high-temperature environment and be multiplied massively in the organic solid wastes; a pile is high in heating speed and temperature, can be quickly composted, can degrade organic matter more quickly, minimization and harmlessness are realized more completely, and the organic solid wastes can be converted into biological fertilizers which contain massive bacillus licheniformis, can inhibit plant pathogens, can promote growth of plants and can improve the yield of crops. The bacillus licheniformis UTM102 has good strain performance and remarkable economic efficiency.

Description

Technical field [0001] The present invention relates to the field of fermentation, in particular to a Bacillus thermophilus strain UTM102 carrying a phytase gene and its use in degrading organic solid waste and preparing biological fertilizers. Background technique [0002] A large amount of organic solid waste is generated in the process of urban and rural life and production. For example, a large amount of sludge is produced in the process of urban domestic sewage treatment, which contains a large number of harmful bacteria, and is rich in organic matter, which is easy to decompose and stink, causing serious secondary pollution. The waste from the production process of agricultural and animal husbandry, such as orange stalks, animal manure and animal carcasses, has become a serious source of pollution, and has become the first source of pollution in some areas. [0003] The main component of urban and rural organic solid waste is lignocellulosic material. Lignocellulose is a cr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C05F17/00A01N63/00A01P21/00C12R1/10
CPCA01N63/00C05F11/08C12N1/20C12N1/205C12R2001/10Y02W30/40Y02A40/10
Inventor 刘永跃何璧梅许宜北汪涌
Owner 大地绿源环保科技(北京)有限公司
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