Bacillus licheniformis with biomass hydrolase activity and application thereof

A technology of bacillus licheniformis and hydrolase, applied in the direction of microorganism-based methods, applications, microorganisms, etc., can solve the problems of incomplete transformation of humus, long fermentation time, etc., and achieve good economic and social benefits and excellent performance

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

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Problems solved by technology

The traditional composting method has problems such as l

Method used

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  • Bacillus licheniformis with biomass hydrolase activity and application thereof
  • Bacillus licheniformis with biomass hydrolase activity and application thereof
  • Bacillus licheniformis with biomass hydrolase activity and application thereof

Examples

Experimental program
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Example Embodiment

[0035] Example 1 Isolation and Identification of Bacillus licheniformis UTM108

[0036] Take about 1g of bottom mud sample of Tengchong Hot Spring in Yunnan Province and place it in a 250ml triangular flask filled with several small glass beads, which contains 100ml of sterile water. Shake on a constant temperature shaker at 40°C for 1 hour and then let stand for 30 minutes. Aseptically draw 1ml of the supernatant and transfer to 100ml of sterilized enrichment medium ((NH 4 ) 2 SO 4 0.06%, K 2 HPO 4 0.2%, MgSO 4 ·7H 2 O 0.04%, K 2 SO 4 0.02%, NaCl 0.01%, CaCl 2 2H 2O 0.03%, hemicellulose 1.5%, agar 1.5%, pH 6.5). Shake enrichment and acclimation culture on a constant temperature shaker at 40°C for 3 days, with a rotation speed of 160 rpm. Take 1ml of the bacterial suspension after 3 days of cultivation, add it to a test tube containing 9ml of sterile water, and make 10ml by gradient dilution method. -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 the dilution. ...

Example Embodiment

[0038] Example 2 Detection of xylan deacetylase activity of bacterial strain UTM108 and its gene sequence

[0039] Take 100ml of UTM108 bacteria liquid (5g of beef extract, 5g of yeast extract, 10g of peptone, 5g of sodium chloride, 5g of hemicellulose, 15g of soluble starch, 1000ml of distilled water, pH7.2, cultured at 40°C for 2 days) and centrifuge (6000 rpm / minutes to 10 minutes) to collect the bacteria. The cells were placed in phosphate buffer (pH 7.4), and the cells were disrupted 20 times in a row using an ultrasonic disruptor (600W) with a working time of 5 seconds and an interval of 5 seconds. Centrifuge (7500 rpm, 10 minutes) to obtain the supernatant, that is, to obtain the crude enzyme solution.

[0040] Xylan deacetylase activity was measured with p-nitrophenol acetate as a substrate, and the reaction mixture contained 500 μl p-nitrophenol acetate (5% suspended in 50 mmol / L sodium phosphate buffer, pH 7.0), 450 μl 50 mmol / L L sodium phosphate buffer (pH 7.0) ...

Example Embodiment

[0042] Example 3 Detection of strain UTM108α-L-arabinofuranosidase activity and related gene sequences

[0043] Inoculate the activated UTM108 strain on the LB slant (tryptone 10g, yeast extract 5g, sodium chloride 10g, agar 15g, distilled water 1000ml, pH 7.2) in xylan-containing medium (peptone 4g, oat xylan Sugar 5g, glucose 4g, sodium chloride 5g, distilled water 1000ml, pH7.2), 40 ℃ shaking (160 rpm) for two days, centrifuged (6000 rpm, 10 minutes) to collect the bacteria. The cells were placed in phosphate buffer (pH 7.4), and the cells were disrupted 20 times in a row with an ultrasonic disruptor (power 600W) for 5 seconds with an interval of 5 seconds. Centrifuge (7500 rpm, 10 minutes) to obtain the supernatant to obtain the crude enzyme solution. Arabinosidase activity is determined by the release of p-nitrophenol (pNP) from the substrate p-nitrophenyl-arabinofuranoside (pNPAF). Take 10μl of the enzyme solution to be tested and add it to 0.18ml 0.1mmol / L pH 5.8 buff...

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Abstract

The invention provides a Bacillus licheniformis strain UTM108 with biomass hydrolase activity. The strain is preserved in the CCCCM CGMCC on September 18, 2014 with the preservation number of CGMCC No.9679. The strain UTM108 can utilize sewage sludge of municipal sewage plants, household garbage, kitchen waste, animal carcasses, livestock manure, crop straw as well as organic solid waste such as traditional Chinese medicine dregs, non-hazardous waste fungus residues, food industry waste and the like produced by certain industries for high-temperature fermentative degradation, pollution caused by organic solid waste materials is treated, environment-friendly bio-fertilizers can also be prepared through composting and fermentation by the strain UTM108, and the strain is excellent in performance and has better economic and social benefits.

Description

technical field [0001] The invention relates to the field of organic solid waste treatment, in particular to a bacillus licheniformis with biomass hydrolase activity and application thereof. Background technique [0002] At present, my country's urban and rural areas produce a large amount of organic solid waste every year. In 2005 alone, the statistics of crop straw reached 800 million tons, while the sludge generated by urban domestic sewage treatment reached 80 million tons every year, and the annual growth rate was 5%. At the same time, the organic solid waste generated in the industrial production process has also reached astonishing data, such as a large number of discarded medicinal materials and medicinal residues after extraction and processing of traditional Chinese medicine, and waste from the food processing industry. These large amounts of organic solid waste have not been effectively utilized, which not only wastes precious resources, but also aggravates enviro...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/11C05F17/00C05F7/00C05F15/00C12R1/10
CPCC05F7/00C05F17/00C12N1/20C12N1/205C12R2001/10Y02W30/40Y02A40/20
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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