Bacillussp., microbial agent and applications of Bacillussp. and microbial agent

The technology of Bacillus cereus and microbial inoculum is applied in the application field of preparing ethanol, and can solve the problems of low cellulose degradation ability and the like

Inactive Publication Date: 2013-05-01
AGRI PRODS PROCESSING RES INST CHINESE ACAD OF TROPICAL AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the shortcomings of existing microbial strains with low cellulose degradabilit

Method used

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  • Bacillussp., microbial agent and applications of Bacillussp. and microbial agent
  • Bacillussp., microbial agent and applications of Bacillussp. and microbial agent
  • Bacillussp., microbial agent and applications of Bacillussp. and microbial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Enrichment and isolation of cellulose-degrading strains

[0030] Take 10 g of sludge sample by aseptic operation, put it into a conical flask filled with 90 mL of sterile water and shake it for 30 min to make it fully mixed to form a 10-1 dilution. After clarification, take the supernatant and transfer it to the rich Collect culture medium; directly draw 10ml of the liquid sample to inoculate the enrichment medium, culture it at 30°C and 180 r. Continuous enrichment culture for 20 days.

[0031] The enriched culture solution was made into 10-2, 10-3, 10-4 concentration dilution solution by gradient dilution method. Take 100 μL of 10-2, 10-3, and 10-4 dilutions and spread them on the primary screening medium plate. After culturing at 28°C for 1 to 2 days, duplicate the plate, and use Gram's iodine solution Submerge the parental plate for 3-5 minutes, the result is as follows figure 1 shown.

[0032] The strains with clear transparent circles around the colonies w...

Embodiment 2

[0037] According to the experimental content and experimental methods recorded in the "Common Bacteria System Identification Manual" [4] and "Actinomycete Systematic Principles, Methods and Practice" [5], the screened strains with the preservation number CGMCC No. 6895 were tested. Bacteria identification.

[0038] 1. Morphological analysis

[0039] The observation of the colony morphology of the strain with the preservation number CGMCC No. 6895 was carried out on a nutrient agar medium. After the strain was streaked and inoculated, it was incubated at a constant temperature of 28°C for 48 hours to observe the colony morphology. The colony of the strain with the preservation number CGMCC No. 6895 is white and slightly shiny, with a rough, flat, nearly round surface and irregular edges. The results are as follows: figure 2 shown.

[0040] 2. Identification of 16s rDNA

[0041] PCR amplification and sequencing of 16s rDNA: The total DNA of the strain with the preservation num...

Embodiment 3

[0050] (1) Inoculate 3 parts by weight of Bacillus sp. with the preservation number CGMCC No. 6895 in 100 parts by weight of nutrient broth medium (liquid), culture at 37°C, take samples during the culture process and pass Observation by direct counting method under microscope until the number of viable bacillus was 0.5×10 10 per gram of culture medium.

[0051] (2) Weigh 30g of bagasse (14.06g of cellulose) into a 500ml Erlenmeyer flask, then add the same amount of inorganic nutrient solution (MgSO 4 .7H 2 O 0.5g, KH 2 PO 4 1.0g, Na 2 PO 4 0.2g, MnSO 4 0.035g, CuSO 4 .5H 2 O 0.007g, FeSO 4 .7H 2 O 0.007g, distilled water 1000ml, sterilized).

[0052] (3) The 0.5×10 obtained in step (1) 10 Each gram of culture medium (10 g) was added to the Erlenmeyer flask in step (2). The Erlenmeyer flask was placed in a biochemical incubator at 30°C for static culture for 30 days, and the water content was controlled at about 70%. The results of the cellulose content after 3...

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Abstract

The invention provides Bacillussp. with a preservation number of CGMCC (China General Microbiological Culture Collection Center) No.6895, a microbial agent containing Bacillussp., an application of Bacillussp. or the microbial agent in cellulose degradation, and an application of Bacillussp. or the microbial agent in preparing alcohol by taking cellulose as a raw material. Bacillussp. with the preservation number of CGMCC No.6895 and the microbial agent containing Bacillussp. have very strong degradation capability to cellulose on an alkaline condition, and can be used for preparing alcohol by taking cellulose as the raw material.

Description

technical field [0001] The present invention relates to a kind of bacillus (Bacillus sp.), and a kind of bacillus ( Bacillus cereus ) microbial agents as active ingredients and their applications; more precisely, it involves a Bacillus sp. with a preservation number of CGMCC No. The application of the above-mentioned bacillus or microbial agent in degrading cellulose and in the preparation of ethanol from cellulose. Background technique [0002] In today's world, facing the problems of energy shortage and environmental pollution, countries all over the world are trying to produce and use fuel ethanol. At present, the production of fuel ethanol mainly uses corn and grains as raw materials. However, China has a large number of potential low-cost cellulose production. Material raw materials, cellulose can be degraded into glucose under the action of cellulase, so that it can be used for fermentation to produce ethanol, thereby greatly expanding the production of ethanol and re...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/00C12P7/10C12R1/085
CPCY02E50/16Y02E50/10
Inventor 崔丽虹李积华魏晓奕张雨良付调坤韩冬辉王飞刘义军
Owner AGRI PRODS PROCESSING RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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