Screening and application of bacillus strain for producing cellulase

A technology of cellulase and Bacillus subtilis, applied in the field of bioengineering, can solve problems such as low production efficiency, low cellulase output, and difficult operation, and achieve environmental protection, good generation stability, and simple and easy process operation Effect

Active Publication Date: 2021-07-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the current problem of low cellulase production or low production efficiency and difficult operation, the present invention screened a strain of Bacillus subtilis from animal feces, which can efficiently produce cellulase, rapidly decompose cellulose, and its own The probiotic function can also be beneficial to the health of animal intestines

Method used

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  • Screening and application of bacillus strain for producing cellulase
  • Screening and application of bacillus strain for producing cellulase
  • Screening and application of bacillus strain for producing cellulase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: the screening of bacterial strain

[0041] (1) Screening of Bacillus subtilis producing cellulase

[0042] Preliminary screening process: Take an appropriate amount of animal feces samples, dissolve them in 100mL of sterilized distilled water, put them in a shaker, shake at 30°C and 200r / min for 30min for enrichment culture. Take the supernatant for ten-fold dilution, draw 10 -4 、10 -5 、10 -6 、10 -7 、10 -8They were respectively coated on CMC-Na screening plates and incubated in a 30°C incubator for 24 hours. Since non-cellulase-producing strains do not produce transparent circles, cellulase-producing strains will appear transparent circles. Therefore, the range of potential strains is determined according to whether there is a cellulose-degrading transparent circle;

[0043] Re-screening process: Use the basal medium to isolate and select the strains with potential cellulose degrading ability. Because the ratio of the diameter of the transparent ci...

Embodiment 2

[0055] Embodiment 2: Bacillus subtilis FMME ZK002 shake flask fermentation

[0056] Step 1: Prepare medium

[0057] Carboxymethylcellulose sodium (CMC-Na) screening medium (g / L): CMC-Na 10, (NH 4 ) 2 SO 4 4. Peptone 1, MgSO 4 ·7H 2 O 0.5, KH 2 PO 4 1. Sterilize at 121°C for 15 minutes;

[0058] Solid activation medium (g / L): sodium chloride 5, beef extract 10, peptone 10, agar 20, distilled water 1.0L.

[0059] Liquid seed medium (g / L): sodium chloride 5, beef extract 10, peptone 10, distilled water 1.0L.

[0060] Liquid fermentation medium (g / L): sodium chloride 5, beef extract 10, peptone 10, distilled water 1.0L.

[0061] Step 2: Seed Preparation

[0062] Dip the bacterial suspension of the strain, streak it on the solid medium, cultivate it in a constant temperature incubator at 30°C for about 24 hours to obtain a single colony, pick a single colony and put it into a 100mL conical flask containing 25mL of liquid seed medium, Cultivate overnight at 30° C. in a ...

Embodiment 3

[0065] Embodiment 3: subtilis Bacillus subtilisFMME ZK002 passage stability

[0066] For specific embodiments, referring to Example 2, the screened Bacillus subtilis FMMEZK002 was subcultured for 12 generations, and the strains of each generation were inoculated into the fermentation medium for shake-flask fermentation. After the fermentation, the cellulase activity was measured. After the subculture, the cellulase activity was shown in Table 2, and the enzyme activity was maintained at 108-118.67U / mL, which indicated that Bacillus subtilisFMME ZK002 has high passage stability.

[0067] The cellulase activity of table 2 subculture strain

[0068]

[0069]

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Abstract

The invention discloses screening and application of a bacillus strain for producing cellulase, and belongs to the technical field of bioengineering. According to the invention, a strain of bacillus subtilis is screened from animal waste, the activity of cellulase produced by fermentation of the bacillus subtilis on a shake flask level reaches 118.67 U / mL, and the bacillus subtilis has good passage stability. The method for producing the cellulase by using the bacillus subtilis is simple and feasible in process operation, low in culture medium cost and suitable for industrial production.

Description

technical field [0001] The invention relates to the screening and application of a cellulase-producing bacillus strain, which belongs to the technical field of bioengineering. Background technique [0002] In addition to their own probiotic properties, feed probiotics can also play a positive role in animal digestion and absorption and intestinal health by producing various chemical substances. For example, for the diarrhea caused by animals that are difficult to absorb the crude fiber rich in feed, feeding probiotics can produce cellulase, which converts the crude fiber that animals cannot directly absorb into monosaccharides that are easily absorbed and utilized by animals, and promotes animal absorption. Enrich its nutritional value. Therefore, cellulase has great application in feed. [0003] Compared with traditional microbial metabolic transformation or fermentation optimization methods, the use of forage probiotics to ferment and produce cellulase can not only reduc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/42A23K10/12A23K10/30A23K10/37A23K20/163C12R1/125
CPCC12N1/20C12N9/2437A23K10/12A23K10/30A23K10/37A23K20/163Y02P60/87
Inventor 陈修来赵阔刘立明高聪刘佳郭亮
Owner JIANGNAN UNIV
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