Process for preparing viable bacillus subtilis preparation

A technology of Bacillus subtilis and live bacteria preparations, which is 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 adopted, the initial concentration of the fermentation medium is too high, and the lag period after inoculation is longer.
[0007] During the fermentation process, more by-products are formed, and the concentration of fermentation products cannot increase, which eventually leads to higher fermentation costs;
[0008] 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;
[0009] 4. The use of flocculation/plate-and-frame filtration or centrifugation t...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: (with 5m 3 Fermentation tank as an example)

[0038] 1) Primary seed culture: 2000ml of primary seed culture medium is placed in a 5000ml Erlenmeyer flask, and sterilized by moist heat at 121°C for 15 minutes. Take 10ml of live Bacillus subtilis freeze-dried tube strains and inoculate them in the primary seed medium, and cultivate them at 30°C for 10 hours; the raw materials and dosage of the medium used for primary seed cultivation are: glucose 0.5%, beef extract 0.5%, peptone 3%, sodium chloride 0.3%, medium pH 7.5;

[0039] 2) Secondary seed culture: 150L of secondary seed culture medium is installed in a 400L fermenter, and sterilized by moist heat at 121° C. for 15 minutes. Inoculate 3000ml of cultured primary seeds into secondary seed medium; cultivate at 30°C for 10 hours; the raw materials and dosage of medium used for secondary seed cultivation are: glucose 0.5%, yeast extract 0.5%, peptone 3% , dipotassium hydrogen phosphate 0.02%, sodium chlo...

Embodiment 2

[0046] Embodiment 2, with embodiment 1, the difference is,

[0047] 1) The raw materials and dosage of the medium used for primary seed cultivation are: glucose 0.5%, beef extract 0.5%, peptone 3%, sodium chloride 1%, medium pH 6.5; 8h.

[0048] 2) The raw materials and dosage of medium used for secondary seed cultivation are: glucose 3%, yeast extract 1%, peptone 2.5%, dipotassium hydrogen phosphate 0.05%, sodium chloride 1%, magnesium sulfate 0.1%, medium Adjust the pH to 6.5; sterilize with damp heat at 115°C for 30 minutes, and incubate at 35°C for 8 hours.

[0049] 3) The raw materials and dosage of the medium used for fermentation are: glucose 3%, soybean meal 4%, corn steep liquor 2%, yeast powder 3%, peptone 3%, dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.2%, manganese sulfate 0.002% , the pH of the medium is 6.5; sterilize with damp heat at 115°C for 30min, and culture at 28°C for 36h.

[0050] 4) Feeding: when the fermentation reaches 12 hours, the g...

Embodiment 3

[0054] Embodiment 3, with embodiment 1, the difference is,

[0055] 1) The raw materials and dosage of the medium used for primary seed cultivation are: glucose 1%, beef extract 1.5%, peptone 1.5%, sodium chloride 0.8%, medium pH 7.0; 115°C damp heat sterilization for 30 minutes, 35°C cultivation 8h.

[0056] 2) The raw materials and dosage of the medium used for secondary seed cultivation are: glucose 1%, yeast extract 1.5%, peptone 1.5%, dipotassium hydrogen phosphate 0.03%, sodium chloride 0.8%, magnesium sulfate 0.08%, medium Adjust the pH to 7.0; sterilize with damp heat at 115°C for 30 minutes, and incubate at 35°C for 8 hours.

[0057] 3) The raw materials and dosage of the medium used for fermentation are: glucose 1%, soybean meal 2%, corn steep liquor 2%, yeast powder 2.5%, peptone 2.5%, dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.2%, manganese sulfate 0.002% , the medium pH is 7.0;

[0058] 4) Feeding: Add glucose feeding medium prepared by 30% gluco...

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Abstract

The invention relates to a process for preparing a viable bacillus subtilis preparation. According to the process, after bacillus subtilis undergoes first-stage germ amplification culture and second-stage germ amplification culture, the bacillus subtilis 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 subtilis 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 subtilis preparation. Background technique [0002] Bacillus subtilis (Bacillus subtilis), is a kind of Bacillus genus. Single cell 0.7~0.8×2~3 microns, uniform coloring. No capsule, perinatal flagella, able to move. Gram-positive bacteria, spore 0.6 ~0.9×1.0~1.5 microns, oval to columnar, located in the center of the cell or slightly biased, the cell does not expand after spore formation. The surface of the colony is rough and opaque, stained white or slightly yellow, and often forms wrinkles when growing in liquid medium It is an aerobic bacterium. It can use protein, various sugars and starch to decompose tryptophan to form indole. It is widely used in genetic research, and the synthesis pathway and regulation mechanism of purine nucleotides in this bacterium are relatively clear. Widely distributed in soil and decaying organic matter, it is easy to reproduce in withered grass juice,...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/125
Inventor 饶犇廖先清周荣华刘芳黄大野万文栓刘晓艳曹春霞杨自文
Owner HUBEI BIOPESTICIDE ENG RES CENT
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