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Tibetan pig source bacillus and application thereof

A technology of Bacillus and Bacillus subtilis, applied in the direction of bacteria, enzymes, biochemical equipment and methods, etc., can solve the problems of long production cycle, low cellulase activity, hindering the practical application of cellulase, etc.

Inactive Publication Date: 2013-12-04
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, cellulase preparations have been made from Trichoderma viride, Aspergillius niger, Fusarium sp., Paecilomyces sp. and Penicillium decumbens, etc. enzyme [10] , and obtained considerable economic benefits in the application of industrial production [11] , but at the present stage in the production of cellulase in my country, there are problems such as high complexity of fiber substrates, low cellulase activity, high production costs, and long production cycles, which hinder the practical application of cellulase hydrolysis. [12] , mainly because Trichoderma generally lacks a limiting factor β-glucosidase, which can accumulate cellobiose, thereby feedback inhibiting enzyme activity and reducing enzymatic hydrolysis efficiency, which has always been a bottleneck problem hindering its large-scale production and application

Method used

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  • Tibetan pig source bacillus and application thereof
  • Tibetan pig source bacillus and application thereof
  • Tibetan pig source bacillus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1: Separation and screening of BY-2

[0071] 1.1 Source and collection of experimental materials

[0072] The experimental samples were collected from 8-month-old healthy Tibetan pigs in Xi'an Tianxin Tibetan Pig Breeding Base. %, concentrate feed 20% (corn 70%, bran 30%). After the experimental pigs were slaughtered, the abdominal cavity was cut open, the cecum was quickly separated, and about 20 cm (take 2 parts) of the intestinal segment was taken, and the two ends of the intestinal tube were ligated and cut off with strings, put in a sealed plastic bag to vent and seal, and put it in the ice box quickly. Take it back to the lab. One part was stored in a -80°C ultra-low temperature refrigerator for subsequent experiments, and the other was placed in a 4°C refrigerator and quickly used to isolate intestinal bacteria.

[0073] 1.2 Isolation and screening of strains

[0074] 1.2.1 Configuration of medium

[0075] Carboxymethylcellulose sodium selection medi...

Embodiment 2

[0081] Example 2: Identification of BY-2

[0082] 2.1 Morphological features

[0083] The BY-2 strain was cultured in liquid LB medium at 37°C and 220rph for 12 hours with shaking. After Gram staining, it was observed under an optical microscope. It could be seen that the bacteria were rod-shaped and stained purple, which belonged to Gram-positive bacteria ( image 3 ).

[0084] The spore morphology of the bacteria: make a smear of the bacteria liquid cultivated for 24 hours, and use the malachite green staining method to stain the spores. The spores are stained green, and the vegetative bodies are stained red ( Figure 4 ).

[0085] 2.2 Physiological and biochemical identification of BY-2

[0086] According to the methods in "Common Bacteria System Identification Manual" and "Berger's Bacterial Identification Manual", the physiological and biochemical characteristics of the experimental strains were determined. The measurement results showed that the physiological and bio...

Embodiment 3

[0095] Example 3: Analysis of growth characteristics of BY-2

[0096] Streak the screened strains on the LB plate, activate culture at 37°C, pick a single clone and inoculate it into 5ml liquid LB medium, shake and culture to OD at 37°C, 220rpm 600 Between 1.0 and 1.5, inoculate 60 mL of fresh LB medium with 1% inoculum amount, take samples at intervals of 2 hours after 4 hours, and measure the absorbance at 600 nm wavelength by spectrophotometer method to draw the bacterial growth curve until 29 hours.

[0097] While measuring the absorbance value of the bacterial solution, take 2mL of the bacterial solution and shake it evenly, centrifuge at 12000r / min for 2min, discard the supernatant, wash with 0.9% normal saline for 3 times, dry at 80°C until constant weight, and weigh , to obtain 2mL of bacterial liquid containing dry bacterial weight W1, and then converted to dry bacterial weight per liter of fermentation broth: W1×500 (g / L). The measurement results showed that BY-2 be...

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Abstract

The invention discloses a Tibetan pig source bacillus and application thereof. Via bacteria morphology identification, biochemistry identification and molecular identification, the disclosed Tibetan pig source bacillus is named as Bacillus subtilis BY-2 according to classification, and the preservation number of the Tibetan pig source bacillus is CCTCC NO: M2012535. The Bacillus subtilis BY-2 is separated from contents accommodated in the caecum of a Tibetan pig, can generate a higher cellulose clastic enzyme, and has the ability to degrade sodium carboxymethyl cellulose.

Description

technical field [0001] The invention relates to a bacterial strain, which is a bacillus capable of decomposing cellulose isolated from the contents of the cecum of Tibetan pigs, which can be used for the extraction of cellulase to facilitate industrial development; or used in animal production to promote Microecological preparations for pigs and other monogastric animals to digest crude fiber, etc. Background technique [0002] 1 Application Fields and Prospects of Cellulase [0003] Because of its inherent advantages, cellulase is widely used in various industries closely related to people's production and life. Microbial cellulase has a wide range of sources and is easy to obtain. With the development of biotechnology, especially genetic engineering technology, it is relatively easy to produce such cellulase. Therefore, microbial cellulase has been widely used in animal husbandry, feed industry, food industry, papermaking and textile industries and other fields [1][2][3...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/42C12R1/125
Inventor 曹斌云杨伟平王建刚孟凡旭马丽姬生跃彭甲银
Owner NORTHWEST A & F UNIV
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