Cellulase-producing Bacillus and application thereof

A technology of cellulase and bacillus, applied in the field of environmental biology, can solve the problems of difficult biodegradation, achieve the effect of improving composting speed, better ability to degrade cellulose, and shortening heating time

Active Publication Date: 2019-03-01
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, natural cellulose molecules are arranged regularly and aggregated into bundles, making it difficult to biodegrade

Method used

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  • Cellulase-producing Bacillus and application thereof
  • Cellulase-producing Bacillus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Screening of Bacillus thermolactis strain B209

[0052] (1) Mix the remaining sludge from food factory sewage treatment with corn stalks at a mass ratio of 1:0.7:0.8, and inoculate 5% (w / w) summer leaf rot in a certain district of Wuxi City, Jiangsu Province. The water content of the mixture is Composting rate is 55%, C / N ratio is 25, composted after mixing evenly, measure the temperature of the compost at 8:00 am and 6:00 pm every day, when the temperature is higher than 55 ℃, turn the compost once a day, as the compost With the prolongation of the processing time, the temperature of the pile body begins to drop. When the temperature is lower than 40°C, stop turning the pile and take samples;

[0053] (2) Weigh 10 g of compost samples, put them into a Erlenmeyer flask with 90 mL of sterilized water, and vibrate on a shaking table at 120 r / min for 1 h to obtain 10 -1 dilutions, and sequentially made 10 -4 、10 -5 and 10 -6 100 μL of the above three dilution...

Embodiment 2

[0055] Example 2 Identification of Bacillus thermolactis strain B209

[0056] After culturing overnight at 45°C on a nutrient agar medium plate, round, cream-colored, irregular-edged, slightly rough colonies with a diameter of 1-4mm are formed, which are positive for Gram staining by optical microscopy, and appear single or 2 -4 The cells are arranged in a chain, rod-shaped, produce spores, and can use carbohydrates to produce acid.

[0057] 16SrDNA sequencing was used for taxonomic molecular identification, and the sequencing results (1506bp) were compared by Blast in NCBI. It was found that its 16SrDNA base sequence had the highest similarity with Bacillus thermolactis strain, and it was identified as Bacillus thermolactis strain.

[0058] Based on the above morphological characteristics, physiological and biochemical characteristics and genetic characteristics, the cellulase-producing strain can be identified as Bacillus thermomolactis strain, named Bacillus thermomolactis ...

Embodiment 3

[0061] Example 3 Bacillus thermolactis strain B209 strain activation and cultivation

[0062] Bacillus thermolactis strain B209 was inoculated onto nutrient agar medium plate, and cultured at 45°C for 22h. The composition of the nutrient agar medium is 15.0 g of glucose, 5.0 g of yeast extract, and 0.2 g of calcium chloride (with two crystal waters).

[0063] The activated strains were inoculated into a 250 mL shake flask filled with 100 mL of liquid medium, and cultured with shaking at 160 r / min for 22 hours at 45°C. The shake flask medium consists of 15.0 g of glucose, 5.0 g of yeast extract, 0.2 g of calcium chloride (with two crystal waters), 1000 mL of water, and a pH of 7.3.

[0064] Inoculate the shake flask culture solution into a 3L fermenter according to the volume ratio of 3%-10%, the stirring speed is 200-600r / min, the ventilation rate is 0.5-1.0vvm, the dissolved oxygen is greater than 10%, and the temperature is 40°C-60°C Fermentation culture 20-30h. The mediu...

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Abstract

The invention discloses cellulase-producing Bacillus and application thereof and belongs to the technical of environmental biology. Bacillus thermolactis strain B209 has good ability to degrade natural cellulose, can well grow with other strains, and has scientific and reasonable ratio; compost made by using the Bacillus thermolactis strain to carry out piling can be heated for a short time; the high temperature lasts long; cellulose degradation rate is high; compost reaction speed can be effectively increased; harmless effect of the compost can be improved; the Bacillus thermolactis strain has a very promising application prospect.

Description

technical field [0001] The invention relates to a cellulase-producing bacillus and an application thereof, belonging to the technical field of environmental biology. Background technique [0002] According to statistics, there are nearly 20 kinds of crop straws in my country, with an annual output of about 7.0×10 8 t, accounting for about 25% of the world's total straw, of which, rice straw 2.0×10 8 t, corn stalks 2.0×10 8 t, wheat straw 1.0×10 8 t, beans and miscellaneous grain crop straw 1.0×10 8 t, straw vines such as peanuts, potatoes and sugar beets 1.0×10 8 t. With the rapid development of social economy and the increase of population, the total amount of crop straw will increase at an annual rate of 5% to 10%. [0003] The most important component in the plant cell wall is cellulose, which determines the structure of the cell wall. Cellulose is a natural polymer compound composed of glucose residues and cellobiose as the basic linking unit. It is the most abund...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/16C05F15/00C05F17/00C12R1/07C12R1/84C12R1/01
CPCC05F7/00C05F17/00C12N1/16C12N1/20C05F17/80C12N1/205C12R2001/07C05F11/00C05F11/08Y02W30/40
Inventor 李秀芬李月月齐希光王新华任月萍
Owner JIANGNAN UNIV
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