Method for producing beta-mannase by adopting lactic acid bacteria cultured by taking konjaku flour as carbon source

A technology of mannanase and konjac flour, applied in biochemical equipment and methods, glycosylation enzymes, enzymes, etc., can solve the biological safety hazards of mannanase-producing bacteria, low activity of lactic acid bacteria producing mannanase, Long fermentation cycle and other issues, to achieve the effects of no biological safety hazards, expanded application scope, and low production costs

Pending Publication Date: 2017-03-22
蔡河齐
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The present invention aims to solve the problems that the existing fungi that produce mannanase have a long fermentation cycle, the hidden dangers of biological safety in mannanase-producing bac...

Method used

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specific Embodiment approach 1

[0026] Specific embodiment one: present embodiment utilizes konjaku flour as the method for carbon source culture lactic acid bacteria to produce β-mannanase, comprises the following steps:

[0027] 1. Inoculate the Lactobacillus plantarum HDRS1 stored at -80°C on the MRS solid medium plate, and culture it upside down at a constant temperature of 25-32°C for 36-60 hours;

[0028] 2. Pick a single colony and place it in MRS liquid medium, shake it at 25~32℃, 120~180rpm for 24~48 hours;

[0029] 3. Then centrifuge at 5000-9000 rpm for 10-15 minutes, discard the supernatant to collect the bacteria, and adjust the cell concentration to 10 with the blank liquid MRS medium. 8 Individual / mL, as seed solution;

[0030] 4. Insert the seed liquid into the fermentation medium with an inoculation amount of 1% to 3%, and vibrate at 30°C and 140rpm for 36 to 48 hours to obtain a fermentation liquid;

[0031] 5. Centrifuge the fermentation broth at 5000-9000 rpm for 10-15 minutes, discard ...

specific Embodiment approach 2

[0033] Specific embodiment two: the difference between this embodiment and specific embodiment one is: the MRS solid medium formula described in step one is: glucose 1g / 100mL, peptone 1g / 100mL, Tween 80 0.1mL / 100mL, dipotassium hydrogen phosphate 0.2g / 100mL, sodium acetate 0.5g / 100mL, triammonium citrate 0.2g / 100mL, magnesium sulfate 0.02g / 100mL, agar 2g / 100mL and distilled water, pH 6.0-6.5. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0034] Specific embodiment 3: The difference between this embodiment and specific embodiment 1 or 2 is that in step 1, the culture is kept upside down at a constant temperature of 28-31° C. for 40-55 hours. Others are the same as in the first or second embodiment.

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Abstract

The invention provides a method for producing beta-mannase by adopting lactic acid bacteria cultured by taking konjaku flour as the carbon source, relating to a method for producing beta-mannase by adopting lactic acid bacteria, for solving the problems that during the producing of mannase by adopting fungi in the prior art, the fermentation period is long, the bacteria for producing mannase has the potential biological safety hazards, and the activity of the lactic acid bacteria in producing the mannase is low. The method comprises the following steps: 1, inoculating lactobacillus plantarum to an MRS solid medium panel, and carrying out culturing; 2, picking single colonies, and carrying out culturing in MRS liquid medium; 3, carrying out centrifugalization, abandoning the supernatant, collecting thalli, and adjusting the cell concentration, thus obtaining the seed solution; 4, inoculating the seed solution to a fermentation medium, thus obtaining fermentation liquor; and 5, centrifugalizing the fermentation liquor, abandoning the thalli, and taking the supernatant, thus obtaining the mannase crude enzyme. The crude enzyme produced by adopting the method provided by the invention is high in enzyme activity, low in production cost, and short in production period, and the produced bacterial strain has no biological safety hazards. The method is applied to the field of microbial fermentation enzyme production.

Description

technical field [0001] The invention relates to a method for producing β-mannanase by lactic acid bacteria. Background technique [0002] β-mannanase (β-mannanase, referred to as mannanase, EC3.2.1.78), is an endohydrolase that can specifically hydrolyze mannan polysaccharides containing β-1,4-mannosidic bonds. hemicellulase. Mannanase is widely used in industry, especially in the food industry, such as for fruit juice clarification to improve the juice yield; degradation products containing 2-10 residues, that is, oligosaccharides, can promote the growth of probiotics in the human intestinal tract. Microbial mannanase has the advantages of wide sources, high activity, and mild action conditions, and is the most important source for basic research and industrial applications. Mannanase-producing microorganisms are mainly molds in fungi and bacillus in bacteria, but the industrial production scale and application fields are often limited due to long fermentation cycle or po...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12N1/20
CPCC12N9/2494C12N1/20C12Y302/01078
Inventor 蔡河齐
Owner 蔡河齐
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