Process for preparing viable bacillus licheniformis preparation

A technology of Bacillus licheniformis and live bacteria preparations, applied in the direction of bacteria, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of high processing cost, low equipment utilization rate, and product effect decline, and achieve the goal of import wind The effect of mild outlet air temperature is low, the risk of fermentation contamination is reduced, and the utilization rate of equipment is improved

Inactive Publication Date: 2014-12-24
HUBEI BIOPESTICIDE ENG RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. The filling coefficient of the fermentation tank is 60%-70%, and the utilization rate of the equipment is not high;
[0006] 2. Due to the batch fermentation used, the initial concentration of the fermentation medium is too high, the lag period after inoculation is longer, and more by-products are formed during the fermentation process, and the concentration of the fermentation product cannot increase, which eventually leads to higher fermentation costs;
[0007] 3. Flocculation / plate-and-frame filtration or centrifugation technology is generally used for the concentration of fermentation broth. These two technologies generally generate a large amount of wastewater, and the subsequent treatment costs are relatively high;
[0008] 4. The use of flocculation / plate-and-frame filtration or centrifugation technology will also cause the loss of active ingredients in the fermentation supernatant, resulting in a decline in the product effect;
[0009] 5. The inlet temperature of spray drying is generally set at 180°C to 200°C, and the outlet temperature is 80°C to 100°C. Under such a high working temperature, the number of bacteria in some varieties will decrease to a certain extent, which directly affects product quality
[0012] Fermentation and drying and granulation will produce a large amount of waste gas, which often contains volatile organic compounds (VOC), hydrogen sulfide, ammonia, mercaptans and other pollutants, accompanied by unpleasant stench. In the past, fermentation enterprises generally Discharge it directly or simply spray it with water, the two treatment methods will cause serious pollution to the environment

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: (with 5m 3 Fermentation tank as an example)

[0036] 1) First-class seed culture: 2000ml first-class seed medium is placed in a 5000ml triangular flask, sterilized by moist heat at 121°C for 30 minutes, and 10ml of live Bacillus licheniformis freeze-dried tube strains are inoculated in the first-class seed medium, at 28°C Cultivate for 12 hours; no contamination is found in the strain, and the shape of the bacteria is good, and then proceed to the next step. The raw materials and dosage of the medium used are: 0.5% starch, 0.5% beef extract, 3% peptone, 0.5% sodium chloride, and the pH of the medium is 7.5.

[0037] 2) Secondary seed culture: 150L secondary seed medium is installed in a 400L fermenter, sterilized by moist heat at 121°C for 30 minutes, and 3000ml of primary seed is inoculated into the secondary seed medium; cultivated at 28°C for 12h; after inspection , if there is no contamination of the bacteria and the shape of the bacteria is good, pr...

Embodiment 2

[0045] Embodiment 2, with embodiment 1, the difference is,

[0046] 1) The raw materials and dosage of the medium used are: 0.5% starch, 1% beef extract, 2.5% peptone, 0.5% sodium chloride, the pH of the medium is 6.5; sterilize at 115°C for 30min, and inoculate at 30°C for 16h.

[0047] 2) The raw materials and dosage of the medium used are: starch 0.5%, yeast extract 1%, peptone 2%, dipotassium hydrogen phosphate 0.05%, sodium chloride 1%, magnesium sulfate 0.1%, and the pH of the medium is adjusted to 6.5 , 121 ° C damp heat sterilization for 15 minutes, 30 ° C after inoculation for 14 hours.

[0048] 3) The raw materials and consumption of the fermentation medium used are: glucose 1.5%, soybean meal 1.1%, corn steep liquor 1.7%, peptone 3%, dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.2%, manganese sulfate 0.002%, and the pH of the medium is 6.5. Incubate at 32°C for 45h.

[0049] 4) Feeding: when the fermentation reaches 15 hours, the glucose feeding mediu...

Embodiment 3

[0053] Embodiment 3, with embodiment 1, the difference is,

[0054] 1) The raw materials and dosage of the medium used are: 1% starch, 1.5% beef extract, 2% peptone, 0.5% sodium chloride, and the pH of the medium is 6.5.

[0055] 2) The raw materials and dosage of the medium used are: starch 1%, yeast extract 1.5%, peptone 2%, dipotassium hydrogen phosphate 0.02%, sodium chloride 0.5%, magnesium sulfate 0.02%, and the pH of the medium is adjusted to 6.5 .

[0056] 3) The raw materials and dosage of the medium used are: glucose 2%, soybean meal 2%, corn steep liquor 1%, peptone 1%, dipotassium hydrogen phosphate 0.01%, magnesium sulfate 0.1%, manganese sulfate 0.002%, and the pH of the medium is 7.5 . Incubate at 28°C for 50h.

[0057] 4) Feeding: Add glucose feeding medium prepared by 20% glucose and 80% water when fermentation reaches 12 hours, and ferment until 44 hours to end;

[0058] 6) Concentrate the fermented liquid with a plate-type evaporative concentrator: when ...

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PUM

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Abstract

The invention relates to a process for preparing a viable bacillus licheniformis preparation. According to the process, after bacillus licheniformis undergoes first-stage germ amplification culture and second-stage germ amplification culture, the bacillus licheniformis is inoculated to a fermentation culture medium for culture, and the number and conversion ratio of bacilli is increased through feeding a supplement carbon source during fermentation, so that the unit yield is increased remarkably, and the unit cost is reduced; at the end of fermentation, fermentation liquor is concentrated with a vacuum film concentrator, so that soluble synergistic substances, growth promoting substances and bacteriostatic active substances can be effectively recovered from the fermentation liquor; the concentrated fermentation liquor is subjected to drying and granulating by multi-level low-temperature spray-fluidized drying equipment, granules are subjected to sorting, fine powder gathered by a cyclone separator returns and is granulated again, and then, a bacillus licheniformis product, of which the CFU of powder per gram is not less than 2.0*10<11>, is obtained; exhaust gas is treated and is discharged after reaching standard. The process has the advantages that effective substances in the fermentation liquor can be reserved to the maximum, the quality of the product is high, the equipment utilization ratio is high, the retention of biological activity is relatively good, waste gas and waste water are discharged after reaching standard, and large-scale industrial production can be achieved.

Description

technical field [0001] The invention relates to a preparation process of a live bacillus licheniformis preparation. Background technique [0002] Bacillus licheniformis is a Gram-positive thermophilic bacterium commonly found in soil. The bacterium is also found in the feathers of birds, especially ground-dwelling birds such as finches and aquatic birds such as ducks, especially in the breast and back feathers. The optimum temperature for enzyme secretion is 37°C. It may exist in the form of spores to resist harsh environments; under favorable conditions, it can exist in a growing state. [0003] Bacillus licheniformis has a variety of uses, such as promoting the growth of normal physiological anaerobic bacteria in the intestine, adjusting the imbalance of intestinal flora, and restoring intestinal function; and ordinary acute bacillary dysentery, etc., all have obvious curative effects; can produce anti-active substances, and have a unique mechanism of biological oxygen ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/10
Inventor 廖先清周荣华饶犇刘芳陈伟闵勇张先进张光阳杨自文
Owner HUBEI BIOPESTICIDE ENG RES CENT
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