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Bacillus licheniformis for expressing lipase and fermentation enzyme production method by bacillus licheniformis

A technology of Bacillus licheniformis and lipase, applied in the field of microorganisms, can solve the problems of limited species, high production cost, low production efficiency, etc., achieve remarkable low temperature resistance and alkali resistance, reduce fermentation cost, and improve fermentation capacity and production. The effect of efficiency

Active Publication Date: 2021-02-19
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, industrially synthesized lipase has limited types, low production efficiency, high production cost, small optimum reaction temperature and pH range, and is restricted by enzyme activity and applicable conditions, so it cannot meet the needs of industrial production. Screening a high-yielding lipase The Bacillus licheniformis strain of the enzyme is imperative, its extracellular lipase activity is high and has a wide range of applications, and the production cost is low, which is of great significance for expanding the application field of lipase on a large scale

Method used

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  • Bacillus licheniformis for expressing lipase and fermentation enzyme production method by bacillus licheniformis
  • Bacillus licheniformis for expressing lipase and fermentation enzyme production method by bacillus licheniformis
  • Bacillus licheniformis for expressing lipase and fermentation enzyme production method by bacillus licheniformis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Screening method for Bacillus licheniformis NCU CS-5.

[0030] (1) Preparation of experimental materials.

[0031] 1. The isolated source is taken from the camphor tree seeds that have been partially deteriorated during storage after collection and shelling in the laboratory.

[0032] 2. Culture medium.

[0033] Nutrient broth medium (g / L): peptone 10.0, beef extract powder 3.0, sodium chloride 5.0, final pH 7.2±0.2, autoclaved at 121°C for 15 minutes, cooled to room temperature, and set aside.

[0034] Agar broth medium: add agar with a final mass concentration of 15.0-20.0g / L to the nutrient broth medium.

[0035] Preparation of olive oil emulsion: Mix olive oil and 0.1g / mL gum arabic at a ratio of 1:4 (v / v), and emulsify for 10 minutes with a high-speed homogenizer to obtain olive oil emulsion.

[0036] Enrichment medium (g / L): glucose 25.0, soybean peptone 5.0, tryptone 15.0, sodium chloride 5.0, olive oil emulsion 2% (v / v), adjust the pH to 7.2, and ...

Embodiment 2

[0050] Example 2. Enzyme production by fermentation of Bacillus licheniformis NCU CS-5.

[0051] The method for producing enzyme by fermentation of Bacillus licheniformis NCU CS-5 mainly includes the following steps.

[0052] Plate culture: Pick a ring of Bacillus licheniformis NCU CS-5 and inoculate it on the nutrient broth agar plate medium, and culture at a constant temperature of 37°C for 36 hours to obtain first-grade seeds.

[0053] Shake flask culture: Take a ring of the primary seeds and insert them into the seed medium, and cultivate them for 36 hours at 37°C and 200r / min to obtain the secondary seed solution.

[0054] Seed tank culture: The bacteria in the secondary seed solution were inserted into the seed tank medium at a ratio of 40% (w / v) of wet weight inoculum, and cultivated for 12 hours at a constant temperature of 35°C and a rotational speed of 200r / min.

[0055] Fermentation tank culture: the seed liquid in the seed tank was inserted into the culture mediu...

Embodiment 3

[0066] Example 3. The optimum temperature range for lipase.

[0067] Taking the lipase of fermented liquid enzyme activity 32000U / mL produced by the present invention as sample, under the condition of pH value 9.7, measure enzyme activity respectively under different temperature (20-50), the relative enzyme activity change of measuring curve like figure 1 As shown, the optimum temperature range of the enzyme is 30°C-40°C.

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Abstract

The invention relates to bacillus licheniformis for expressing lipase and a fermentation enzyme production method by the bacillus licheniformis, the bacillus licheniformis is preserved in China General Microbiological Culture Collection Center with the preservation number of CGMCC No. 20140. The enzyme activity of fermentation liquor for producing lipase through fermentation of the bacillus licheniformis is as high as 32000U / mL. The optimum action temperature range of the produced lipase is 30 DEG C to 40 DEG C, the optimum pH range is 8.5-9.5, the relative enzyme activity is 60% after heat preservation is conducted at the temperature of 4 DEG C for 48 h, the lipase has wide optimum action range and remarkable low-temperature resistance and alkali resistance, the lipase can be applied to industrial production with different requirements, the industrial application range of the lipase is remarkably expanded, and the application value of the lipase is increased.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a Bacillus licheniformis expressing lipase and a fermentation method thereof. Background technique [0002] Lipase (1ipase, E.C.3.1.1.3), also known as lipolysis, glyceride hydrolase or triacylglycerol acyl hydrolase, can catalyze the hydrolysis of triglycerides to convert them into glycerol and fatty acids at the oil-water interface, Has the property and enantioselectivity to reverse this reaction in both aqueous and non-aqueous media. Lipase has become one of the leading biocatalysts and can be used in many industrial fields such as food, medicine, detergent, textile, biodiesel, paper making, leather, cosmetics and environmental protection. [0003] Lipase mainly comes from animals, plants and microorganisms. Due to the variety of microorganisms, fast reproduction, easy genetic variation, and wider action pH, temperature range and substrate specificity than anima...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/20C12R1/10
CPCC12N1/20C12N9/20C12Y301/01003C12R2001/10C12N1/205
Inventor 曾哲灵赵钧馨曾诚马毛毛余平万冬满
Owner NANCHANG UNIV