Bacillus licheniformis UTM107 producing high-temperature-resistant keratinase and application thereof

A technology of Bacillus licheniformis and UTM107, which is applied in the application, preparation of bacteria, and organic fertilizers, can solve problems such as air pollution, surrounding environment and groundwater pollution, and land occupation, and achieve good social and economic benefits

Active Publication Date: 2015-04-08
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

While consuming a lot of energy and polluting the air, it takes up a lot of land and may also cause pollution to the surrounding environment and groundwater resources.

Method used

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  • Bacillus licheniformis UTM107 producing high-temperature-resistant keratinase and application thereof
  • Bacillus licheniformis UTM107 producing high-temperature-resistant keratinase and application thereof
  • Bacillus licheniformis UTM107 producing high-temperature-resistant keratinase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Isolation and Identification of Bacillus licheniformis UTM107

[0035] Take about 1g of bottom mud sample of Tengchong Hot Spring in Yunnan Province and place it in a 250ml Erlenmeyer flask filled with multiple small glass beads, which contains 100ml of sterile water. Shake on a constant temperature shaker at 50°C for 1 hour and then let stand for 30 minutes. Take 1ml of the supernatant under sterile conditions and connect it to 100ml of sterilized enrichment medium (peptone 1.0%, beef extract 1.0%, casein 1.0%, NaCl 0.5%, K 2 HPO 4 0.07%), cultured with shaking at 50° C. for 3 days at a rotational speed of 160 rpm. Take 1ml of the bacterial suspension after culturing for 3 days, add it to a test tube filled with 9ml sterile water, and prepare 10ml by gradient dilution method. -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 the dilution. Take 0.1ml of each dilution and spread it on the casein selection medium plate (glucose 1.5%, asparagine 0.09%,...

Embodiment 2

[0038] Example 2 Detection of high temperature resistant keratinase activity and its gene sequence of bacterial strain UTM107

[0039] The UTM107 bacterial strain obtained in Example 1 was inoculated in solid wool-containing medium (glucose 1.5%, beef extract 0.1%, wool powder 0.2%, asparagine 0.11%, K 2 HPO 4 0.12%, agar 1.5%, pH 6.4) plate and casein medium (glucose 1.5%, beef extract 0.07%, casein 2.0%, asparagine 0.09%, K 2 HPO 4 0.07%, agar 1.5%), cultured at 50°C for 3 days. Observe that it all has transparent hydrolysis circle on above-mentioned two kinds of medium, confirms that UTM107 bacterial strain can decompose keratinase, and it has keratinase activity. UTM107 bacterial strain is inoculated into liquid medium (1% feather meal, 0.1% dipotassium hydrogen phosphate , 0.01% MgCl 2 ; Prepared with tap water, and adjusted the pH value to 7.0-8.0 after the preparation), cultivated at 50°C for 48 hours, centrifuged at 5000 rpm, took 1.0ml supernatant, and added 2.0ml...

Embodiment 3

[0041] Example 3 Detection of Allantoinase Activity of Bacterial Strain UTM107 and Related Gene Sequences

[0042] UTM107 in the fermentation medium (peptone 1.0%, beef extract 1.0%, casein 1.0%, NaCl 0.5%, K 2 HPO 4 0.07%) at 40°C for 2 days, centrifuged at 4000 rpm to take the supernatant as a crude enzyme solution, and rapidly measured its enzyme activity. The method for measuring allantoinase activity is: add 100 μl of UTM107 crude enzyme solution (supernatant after fermentation and centrifugation) to 2ml of 50mmol / L Tris-HCl buffer (pH 7.5) containing 15mmol / L allantoin, 30 The reaction was carried out at ℃ for 15 minutes, and then a drop of concentrated hydrochloric acid was added to terminate the reaction. Take 0.5ml of reaction solution to measure the content of allantoic acid in the enzyme reaction product to measure the activity of allantoinase. The determination of allantoic acid adopts high performance liquid chromatography detection method, adopts C 18 The col...

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Abstract

The invention provides bacillus licheniformis (Bacillus licheniformis) UTM107 producing high-temperature-resistant keratinase and application thereof. The strain has the biological collection number of CGMCC No.9678. The UTM107 strain can be used for aerobic composting fermentation in downflow sludge, household refuse, kitchen wastes, carcass, excrements of livestocks and other wastes, the temperature can reach 80-90 DEG C in the fermentation process, the highest temperature can be over 100 DEG C, the high temperature can be maintained for a long time, and organic matters in the wastes can be effectively decomposed. The bio-fertilizer prepared by the strain disclosed by the invention is environment-friendly and excellent in performance, and harmlessness, reduction and recycling can be realized when the UTM107 strain is used for treating the organic solid wastes. The bio-fertilizer prepared by the invention is environment-friendly and excellent in performance.

Description

technical field [0001] The invention relates to the field of urban and rural organic solid waste treatment, and relates to a Bacillus licheniformis UTM107 strain containing keratinase gene and its application in urban and rural organic solid waste treatment. Background technique [0002] The domestic sludge produced in cities and towns in my country is about 80 million tons every year, and it is increasing at a rate of 5%. A large amount of sludge is produced after urban domestic wastewater is treated, and its treatment method is mainly landfill after dehydration. While consuming a lot of energy and polluting the air, it takes up a lot of land and may also cause pollution of the surrounding environment and groundwater resources. In addition, the production capacity of my country's livestock and poultry breeding industry ranks first in the world at present, and a large amount of organic waste generated in the process of livestock and poultry breeding and processing has becom...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C05F17/00C12R1/10
CPCC05F17/80C12N1/205C12R2001/10Y02W30/40
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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